Expert Systems With Applications 202 (2022) 117238

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Expert Systems With Applications

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Review

Classification of non-Hodgkin lymphomas based on sample entropy

signatures
Guilherme Botazzo Rozendo a ,∗, Marcelo Zanchetta do Nascimento b , Guilherme Freire Roberto b ,
Paulo Rogério de Faria d , Adriano Barbosa Silva b , Thaína Aparecida Azevedo Tosta c , Leandro
Alves Neves a
a
Department of Computer Science and Statistics (DCCE), São Paulo State University (UNESP), Rua Cristóvão Colombo, 2265, 15054-000, São José do Rio Preto,
São Paulo, Brazil
b
Faculty of Computer Science (FACOM) - Federal University of Uberlândia (UFU), Avenida João Naves de Ávila 2121, Bl.B, 38400-902, Uberlândia, Minas
Gerais, Brazil
c
Science and Technology Institute, Federal University of São Paulo (UNIFESP), Avenida Cesare Mansueto Giulio Lattes, 1201, 12247-014, São José dos Campos,
São Paulo, Brazil
d Department of Histology and Morphology, Institute of Biomedical Science, Federal University of Uberlândia (UFU), Av. Amazonas,

S/N, 38405-320, Uberlândia, MG, Brazil

ARTICLE INFO ABSTRACT

Keywords: Computational systems to provide studies and diagnoses of non-Hodgkin’s lymphomas have been increasingly
SampEn signatures developed to assist specialists in their decision-making. On the other hand, the approaches have not yet
Signature metrics fully explored the sample entropy to assess the disease’s characteristics, with indications of the best features
Non-Hodgkin’s lymphomas
and techniques to distinguish the main groups of lymphomas. Herein, we present a method that considers
Segmentation
sample entropy signatures to study chronic lymphocytic leukemia, follicular lymphoma, and mantle cell
Color normalization
lymphoma, involving images after applying multiple segmentation techniques and color normalization models.
Texture signatures were defined as feature curves obtained from multiple observations, by associating different
parameters. Each signature was calculated for windows of size 𝑚 and ranged from 1 to 4 (0.06–0.40)
tolerance levels (𝑟). The behavior of each signature was determined from the area under the curve, skewness,
maximum entropy value and area ratio, representing the signature metrics. This approach aimed to improve
the quantitative ability of this descriptor, with new interpretations and associations. Thus, all features were
provided for different functions, such as lazy learning, trees, genetic evolution, particle swarm and animal
behavior-based approaches. The best association was tested after adding noise levels of 20%, 40% and 80%.
The performances achieved with the proposal were accuracy rates between 98.72% and 99.60%. We were
able to obtain an accuracy of 98.84% for multi-class classification using only ten features, which is an amount
considerably lower than most of the state-of-the-art approaches. The proposed approach presents relevant
advances for the investigation of computer-aided diagnosis systems since it provides accurate results and
does not require preprocessing steps. Moreover, signature metrics in association with evolutionary approaches,
which provided the best results, are among the main contributions of this paper, since the details of the most
relevant features with a full understanding of the solutions are useful for the study and pattern recognition of
non-Hodgkin’s lymphomas.

1. Introduction

Non-Hodgkin’s lymphomas (NHL) are malignant tumors that affect

cells of the lymphatic system. The American Cancer Society estimates

The code (and data) in this article has been certified as Reproducible by Code Ocean: (https://codeocean.com/). More information on the Reproducibility
Badge Initiative is available at https://www.elsevier.com/physical-sciences-and-engineering/computer-science/journals.
∗ Corresponding author.
E-mail addresses: guilherme.botazzo@unesp.br (G.B. Rozendo), marcelo.zanchetta@gmail.com (M.Z. do Nascimento), guilhermefroberto@gmail.com
(G.F. Roberto), paulo.faria@ufu.br (P.R. de Faria), adrianobs@gmail.com (A.B. Silva), tosta.thaina@gmail.com (T.A.A. Tosta), leandro.neves@unesp.br
(L.A. Neves).

https://doi.org/10.1016/j.eswa.2022.117238
Received 18 January 2021; Received in revised form 6 April 2022; Accepted 10 April 2022
Available online 19 April 2022
0957-4174/© 2022 Elsevier Ltd. All rights reserved.
G.B. Rozendo et al.
that 80,470 new cases of NHL will be diagnosed in 2022 and 20,250
NHL deaths are estimated in the United States (American Cancer Soci-
ety, 2022a). In Brazil, 36,090 new cases of NHL are expected to be
diagnosed throughout the three-year period spanning from 2020 to
2022 (Instituto Nacional de Câncer, 2022).
The World Health Organization classifies NHL according to lym-
phocyte types and histological characteristics of tissues affected by
the disease, in addition to chromosomal information and the presence
of specific proteins within cell membranes (American Cancer Society,
2022b; Swerdlow et al., 2016). Among NHL types, chronic lymphocytic
leukemia (CLL), follicular lymphoma (FL) and mantle cell lymphoma
(MCL) are particularly important. CLL is characterized by slow prolif-
eration of lymph node B and 𝑇 cells, which are present in the blood and
the bone marrow (American Cancer Society, 2022b; Hallek, 2017). FL
occurs both in lymph nodes and bone marrow, which is composed of
two types of B-cells derived from its germinal center: centrocytes and
centroblasts (American Cancer Society, 2022b; Xerri et al., 2016). The
MCL group proliferates throughout the lymph nodes, spleen, blood and
bone marrow (Vose, 2017).
The diagnosis of the NHL type is essential to define the most appro-
priate treatment protocol for each patient. Diagnosis can be performed
via a biopsy of the most accessible tumor. The confirmation of a NHL
type can be accomplished via microscopic evaluation of hematoxylin
and eosin (H&E) stained samples by pathologists. Visual analysis is
a complex task due to the significant time involved, its subjectivity
and pathological interpretation variability between specialists during
the manual diagnosis stage (Albayrak et al., 2021; Ramachandran,
Strisciuglio, Vinekar, John, & Azzopardi, 2020). Computational vision
algorithms and image processing techniques employed in the computer-
aided diagnosis system (CAD) contribute to the diagnosis stage of
digitized images of histological samples of NHL (Bai, Jiang, Li, & Ma,
2019; Battula & Sharma, 2018; Brancati, De Pietro, Frucci, & Riccio,
2019; Candelero et al., 2020; do Nascimento, Martins, Tosta, & Neves,
2018; Jiang, Li, Li, & Zhou, 2018; Li et al., 2022; Martins et al., 2019;
Ribeiro et al., 2018; Roberto, Lumini, Neves, & do Nascimento, 2021;
Taino et al., 2020; Tambe, Mahajan, Shah, Agrawal, & Garware, 2019;
Zhang, Zhang, Jiang, & Yang, 2021).
A CAD system can consider different stages, such as image prepro-
cessing, segmentation, feature extraction and classification, in order to
allow greater objectivity and less variability in the process of image
analysis. Each stage brings important challenges for the appropriate
development of this type of system. For instance, NHL images obtained
from the biopsy sample may show non-uniform staining due to the
H&E process (Feng, Zhang, & Yi, 2018; Jothi & Rajam, 2017). Thus,
this problem can be minimized during the preprocessing stage by
applying color normalization techniques (Khan, Rajpoot, Treanor, &
Magee, 2014; Macenko et al., 2009; Reinhard, Adhikhmin, Gooch, &
Shirley, 2001; Vahadane et al., 2016). This preprocessing step can
slightly increase the average accuracy of texture classification (Cer-
nadas, Fernández-Delgado, González-Rufino, & Carrión, 2017). Also,
segmentation approaches are commonly applied to identify the region
of interest in lymphoma images (De Oliveira et al., 2013; Silva et al.,
2019; Tang, Li, & Xu, 2018; Tokime, Elassady, & Akhloufi, 2018; Tosta,
Faria, Neves, & do Nascimento, 2017; Vahadane & Sethi, 2013; Wienert
et al., 2012), such as lymphocytes and cell nuclei, in order to allow a
correlation with the disease under investigation. In this context, a study
of the most appropriate approaches, capable of providing accurate re-
sults for the improvement of subsequent stages, especially the features
extraction and classification steps, comprise an important challenge in
the study of NHL images. In addition, a model considering a reduced
number of features can lead to a more comprehensive solution for the
community interested in the study of NHL.
In order to present feasible solutions to the aforementioned chal-
lenges, feature extraction and classification stages comprise an impor-
tant part of the CAD systems. These steps are useful for associating
the intrinsic properties of the regions of interest with the pathologies,
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Expert Systems With Applications 202 (2022) 117238
providing solutions with a more complete understanding of the classifi-
cation process, as well as a faster and more accurate diagnosis (Ribeiro
et al., 2018). In this context, investigations have contributed to the
improvement of CAD systems devoted to NHL images and supporting
specialists in their decision-making (Bai et al., 2019; Battula & Sharma,
2018; Brancati et al., 2019; Candelero et al., 2020; do Nascimento
et al., 2018; Jiang et al., 2018; Martins et al., 2019; Ribeiro et al.,
2018; Roberto et al., 2021; Taino et al., 2020; Tambe et al., 2019).
These methods have used different combinations of feature descriptors
and classification techniques, such as Fourier and wavelet transforms,
multiscale and multidimensional approaches, morphological and non-
morphological models, fractal analysis and deep learning strategies.
Part of these approaches explores texture which is determined by the
spatial arrangement of the intensity levels of pixels.
Regarding the importance of the previously mentioned descrip-
tors, features based on sample entropy (SampEn) have not been fully
explored to assess NHL images (Humeau-Heurtier, 2019), although en-
tropy models are considered relevant for quantifying patterns in digital
images (Azami, da Silva, Omoto, & Humeau-Heurtier, 2019; da Silva,
da Silva Senra Filho, Fazan, Felipe, & Murta, 2014; dos Santos et al.,
2018; Fabbri, Goncalves, Lopes, & Bruno, 2012; Gaudêncio et al., 2020;
Haralick, Shanmugam, et al., 1973; Humeau-Heurtier, 2019, 2021;
Morel & Humeau-Heurtier, 2021; Silva, Senra Filho, Fazan, Felipe, &
Junior, 2016; Yeh, Lin, & Shieh, 2011). In addition, this descriptor can
be applied to analyze and classify NHL images by providing a measure
of disorder of the histological structures that represent the groups under
investigation, considering multiple segmentation and color normaliza-
tion techniques. In ordered systems, for instance, entropy is equal to
zero, and as the degree of disorder increases, entropy also increases. In
digital images, the disorder is related to texture (Azami et al., 2019;
Humeau-Heurtier, 2019, 2021; Morel & Humeau-Heurtier, 2021). This
type of investigation has not been fully detailed in the context of NHL
images, with the indications of the main contributions and limitations
of the SampEn for distinguishing the lymphoma groups.
Studies have shown that there are several challenges associated with
the use of SampEn for quantifying and classifying patterns. The capacity
of SampEn depends mostly on establishing the best combinations of its
parameters, especially image sampling windows (sized 𝑚×𝑚) and a tol-
erance level (𝑟) in order to indicate the differences between all pixels of
two windows (da Silva Senra Filho et al., 2019; dos Santos et al., 2018;
Humeau-Heurtier, 2019; Richman & Moorman, 2000; Silva, Duque,
Felipe, Murta, & Humeau-Heurtier, 2018; Silva et al., 2016; Yentes
et al., 2013). This difficulty can be minimized by using the parameters
𝑚 and 𝑟 to define texture signatures based on SampEn, a strategy that
has not been entirely explored with this type of entropy, especially
when NHL images are considered (Li et al., 2022). This approach avoids
the need to indicate a single combination of 𝑚 and 𝑟 for texture analysis,
in addition to expanding the quantitative capacity of SampEn and en-
couraging new interpretations for this descriptor. It is noticed that the
concept of texture signatures has been used and interpreted in different
ways, mainly in the context of fractal techniques (Backes & Bruno,
2013; Backes, Casanova, & Bruno, 2012; Căliman & Ivanovici, 2012;
Dong, 2008; Peleg, Naor, Hartley, & Avnir, 1984). Texture signatures
are useful for organizing multiple quantifications as feature curves. This
strategy can contribute to the pattern recognition of complex medical
images, such as for distinguishing the lymphoma groups. Moreover,
the discriminative power of SampEn signatures has not been verified,
especially considering the different modules that support the devel-
opment of CAD system, such as normalization strategies (Khan et al.,
2014; Macenko et al., 2009; Reinhard et al., 2001; Vahadane et al.,
2016), segmentation techniques (De Oliveira et al., 2013; Silva et al.,
2019; Tang et al., 2018; Tokime et al., 2018; Tosta, Faria, et al., 2017;
Vahadane & Sethi, 2013; Wienert et al., 2012) and tests with distinct
and elevated types of noise (Feng et al., 2018; Zhu & Wu, 2004). These
advances may encourage the study of new solutions for each stage of a
CAD system, both for NHL samples and other contexts of H&E images.
G.B. Rozendo et al.
In this paper, a method based on SampEn signatures and association
with different machine learning algorithms is proposed to study the
NHL cancer groups, involving images after applying multiple segmen-
tation techniques and color normalization models. SampEn signatures
consisted in feature curves wherein each point represents a quantifica-
tion obtained through the execution of SampEn with a configuration
of the parameters 𝑚 and 𝑟. The values for 𝑚 and 𝑟 were evaluated in
the range from 1 to 4 and 0.06 to 0.4, respectively. Each signature
was analyzed with the area under the curve, skewness, maximum
entropy and area ratios attributes, representing the signature metrics,
a strategy that had not been explored in the literature. The attributes
were employed to elaborate the feature vectors which were provided
as inputs for distinct classifiers. The model was tested by considering
NHL images obtained from segmentation techniques (De Oliveira et al.,
2013; Silva et al., 2019; Tang et al., 2018; Tokime et al., 2018; Tosta,
Faria, et al., 2017; Vahadane & Sethi, 2013; Wienert et al., 2012) and
color normalization approaches (Khan et al., 2014; Macenko et al.,
2009; Reinhard et al., 2001; Vahadane et al., 2016). Finally, the dis-
criminative power of the best association, using a reduced number of
features, was evaluated under some conditions (after applying different
types of noise, in addition to comparisons with previous works and
other texture attributes).
This approach will contribute to the diagnosis stage of NHL histo-
logical images, providing the following assessments:
1. A detailed experimental analysis dedicated to improving tech-
niques for CAD systems on NHL images including color nor-
malization, segmentation, feature extraction and classification
approaches;
2. Association of SampEn signatures and different classifiers for
evaluating H&E images from an important dataset that is com-
monly investigated in the literature;
3. Evaluation of the discriminative power of SampEn signatures on
histological lymphoma images, involving images obtained after
applying multiple segmentation techniques and color normaliza-
tion models;
4. Enhancement of the quantitative ability of sample entropy with
evaluations based on signature metrics;
5. Identification of the best SampEn signature metric such as area
under the curve, skewness, maximum point and area ratio for
achieving pattern recognition and discriminative power with
regard to CLL, FL and MCL groups;
6. Definition of an approach that best separates the NHL cancer
groups, even when tests with elevated noise levels are applied
to the signature metrics attributes.
This paper is organized as follows. Section 2 presents an overview of
the works related to our proposal. Section 3 describes the methodology
for the development of the model. Section 4 indicates the tests and the
results achieved in each combination. A detailed discussion is presented
in Section 5, considering the best combination and different conditions.
Finally, we conclude the paper in Section 6, with an overview of the
obtained results and suggestions for future researches.
2. Related work: An overview
Although the CAD systems have been presented to support special-
ists in their decision-making regarding NHL groups, the application of
SampEn signatures, signature metrics and combination with different
machine learning algorithms, commonly used in the different modules
of a CAD strategy, have not been explored in the literature.
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Expert Systems With Applications 202 (2022) 117238
2.1. Sample entropy
Different approaches have been presented to explore entropy in
different contexts (Gaudêncio et al., 2020; Humeau-Heurtier, 2019,
2021; Morel & Humeau-Heurtier, 2021). One such technique is named
sample entropy (SampEn). SampEn was designed to measure irregu-
larity in short and noisy temporal series (Song, Crowcroft, & Zhang,
2012). This approach was applied to assess one-dimensional signal
studies, such as heartbeat variability (Costa, Goldberger, & Peng, 2002),
electroencephalogram data (Song et al., 2012) and atrial fibrillation via
electrocardiogram (Alcaraz & Rieta, 2010). By taking into consideration
these studies, the SampEn was able to quantify the complexity, or irreg-
ularity, of a signal by sampling of similar standards contained therein.
Moreover, the irregularity was inherently related to unpredictability.
Thus, for regular signals, patterns are predictable, and consequently,
entropy is close to zero. On the other hand, the unpredictability in-
creases with the irregularity and, therefore, the entropy also increases.
It is important to notice that the SampEn concept has been expanded
and applied to images (da Silva et al., 2014; da Silva Senra Filho et al.,
2019; dos Santos et al., 2018; Humeau-Heurtier, 2019, 2021; Silva
et al., 2018, 2016; Yeh et al., 2011). However, SampEn signatures and
signature metrics were not explored in these works (Li et al., 2022).
Then, this study has evaluated these metrics to investigate texture
irregularities applied in NHL images.
Regarding two-dimensional SampEn, texture irregularity can be
calculated from image sampling windows (sized 𝑚 × 𝑚), which are used
to measure the similarity of evaluated windows (Yeh et al., 2011). This
method is performed by setting a window at a point on an image and
comparing correspondent pixels within a sliding window that scans
the whole image by incrementally adjusting its position by one unit.
If the differences observed between all pixels of two windows are
smaller than a tolerance parameter 𝑟, then the windows are considered
similar. SampEn is calculated by determining the logarithmic ratio of
the average quantity of similar patterns observed in windows 𝑚 + 1
and 𝑚. If the average quantity of similar patterns in 𝑚 + 1 is equal to
the average amount determined for 𝑚, then the patterns are considered
predictable and the SampEn value is defined as zero. As the number
of similar patterns decreases, the predictability of the patterns also
decreases and SampEn values increase. This type of image verification
shows that if the analyzed texture is uniform, the subsequent state
(𝑚 + 1) will be predictable in relation to the previous state (𝑚), and
consequently, entropy is close to zero. This process can lead to a single
combination of 𝑚 and 𝑟 for texture analysis and limit the quantification
and interpretation of complex images (da Silva et al., 2014; da Silva
Senra Filho et al., 2019; dos Santos et al., 2018; Humeau-Heurtier,
2019, 2021; Silva et al., 2018, 2016; Yeh et al., 2011). On the other
hand, the application of SampEn in the texture analysis of digital
images has advantages, mainly because the process of pattern sampling
using a sliding window is useful for quantifying the spatial distribution
of pixels, a characteristic that is not present in methods based on
histograms, such as Shannon’s entropy approach (Wibmer et al., 2015).
In order to improve and explore the discriminative power of Sam-
pEn, different strategies have been presented. For instance, SampEn can
be used for separating the epithelial nuclei from the stroma in H&E
stained images (Ali et al., 2020), in addition to presenting relevant
results in different contexts.
Yeh et al. (2011) proposed an extension of the multiscale Sam-
pEn of Costa et al. (2002) for the context of digital images. In their
method, called multiscale complexity, the coarse-grained process was
used as a multiscale strategy. The authors proposed a strategy using
a subset of randomly selected windows for computing SampEn faster.
SampEn was computed on each scale and, the scale that provided
the highest differentiation between lymphoma classes was determined
using the ANOVA and the Student–Newman–Keuls test. The authors
concluded that SampEn is a relevant technique for complex medical
image analysis.
G.B. Rozendo et al.
Both da Silva et al. (2014) and Silva et al. (2016) applied SampEn
in rat sural nerve images. The authors explored several values for the
SampEn parameters and, the AUC was used as a criterion to select the
combination of parameters that provided the highest power discrimi-
nation. The authors indicated that SampEn is robust for the analysis
of small images. It can be useful in histological image studies, mainly
because it was possible to detect different properties of the image that
were not directly related to the morphometry of the objects.
dos Santos et al. (2018) proposed a multidimensional and multiscale
version of the SampEn associated with the fuzzy strategy to classify col-
orectal cancer images. The technique considered different window sizes
and different tolerance values to implement the multiscale approach.
The multidimensional approach was implemented considering each
image pixel as a 𝑛-dimensional color vector and the fuzzy approach was
used to evaluate the relevance of the distances between the pixels of
the windows. The authors concluded that this association of techniques
with the logistic classifier allowed an AUC of 0.983, an important rate
given the difficulties of clinical practice for the diagnosis of this type
of cancer.
Raju, Pabboju, and Rao (2020) combined the Dual-Tree M-Band
wavelet transform with SampEn to extract features in brain MRI im-
ages. The authors aimed to separate the images between normal and
abnormal categories. The Naïve Bayes classifier was used. The method
provided 97% accuracy, 98% specificity and 96% sensitivity.
Despite the advances presented previously, the discriminative abil-
ity of SampEn to quantify distinct patterns depends on establishing
the best combinations of its parameters, such as the parameters 𝑚 and
𝑟 (da Silva Senra Filho et al., 2019; dos Santos et al., 2018; Humeau-
Heurtier, 2019; Richman & Moorman, 2000; Silva et al., 2018, 2016;
Yentes et al., 2013). A possible solution considers the strategy based
on texture signature, which has been widely applied in the context of
fractal techniques (Backes & Bruno, 2013; Backes et al., 2012; Căliman
& Ivanovici, 2012; Dong, 2008; Peleg et al., 1984; Roberto et al.,
2017). For instance, Backes et al. (2012) combined signatures with
the fractal dimension descriptor. The authors indicated that signatures
provided a richness of image texture detail, which allowed more ac-
curate quantification. The signatures from images were also applied
in association with neural networks (Junior, Backes, & Bruno, 2018).
The signatures were used to describe the color textures of the images.
The authors performed tests on several image databases and, the results
were promising. The classification performance of the proposed tech-
nique outperformed most of the methods compared, indicating that the
technique can be applied in different contexts. Thus, when the SampEn
is considered, texture signatures can be defined through the parameters
𝑚 and 𝑟 in order to improve the quantitative capacity of this descriptor,
with new interpretations.
These indications have motivated us to investigate a combination
able to improve the classification of NHL images with tests performed
under different conditions. This strategy has not been fully applied as
in our proposal (Li et al., 2022), mainly with the analysis of tech-
niques present in different modules of a CAD system: normalization
approaches (Khan et al., 2014; Macenko et al., 2009; Reinhard et al.,
2001; Vahadane et al., 2016); segmentation techniques (De Oliveira
et al., 2013; Silva et al., 2019; Tang et al., 2018; Tokime et al., 2018;
Tosta, Faria, et al., 2017; Vahadane & Sethi, 2013; Wienert et al., 2012);
distinct and elevated types of noise (Feng et al., 2018; Zhu & Wu, 2004).
2.2. NHL classification systems
Methods for CAD systems have been developed to achieve the best
distinctions in the NHL categories, using multiple techniques (Li et al.,
2022).
Orlov et al. (2010) applied Fourier and Wavelet transforms as
feature quantifiers in NHL images. According to the authors, the quan-
tification performed in the H&E color space with the weighted neighbor
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Expert Systems With Applications 202 (2022) 117238
distance using compound hierarchy of algorithms representing mor-
phology (WND-CHARM) classifier provided the best result, which was
99% accurate. Nascimento, Neves, Duarte, Duarte, and Batista (2015)
performed the classification of NHL classes using the stationary wavelet
transform as a characteristic quantifier. The authors used the ANOVA
method to remove possible noise and the support vector machine (SVM)
classifier, obtaining 100% of accuracy.
Color, texture and visual descriptors were also explored in the liter-
ature for the investigation of NHL cancer. Meng, Lin, Shyu, and Chen
(2010) developed a quantification strategy that consisted of dividing
each NHL image into 25 sub-images and extracting 12,625 features
related to color and texture from each sub-image. The chi-square
feature selection method was applied to select the 50 most relevant
attributes and, the accuracy obtained was 92.70%. Song, Cai, Huang,
Feng, Wang, and Chen (2016) proposed a method capable of extracting
visual attributes from grayscale NHL images and analyzing the results
from different classifiers. The best result was accuracy of 96.80%
with the SVM classifier. Codella, Moradi, Matasar, Sveda-Mahmood,
and Smith (2016) presented a technique to obtain twelve descriptors
from segmented NHL images, considering different spatial and color
configurations. The authors also extracted visual descriptors from the
images. The SVM classifier was applied and the result was an accuracy
rate of 95.50%.
In addition to the techniques previously presented, Roberto et al.
(2017) described a method based on percolation theory combined
with multiscale and multidimensional strategies. The method analyzed
different combinations of features, segmentation methods and, classi-
fiers. The best result was an accuracy rate of 96.40%. do Nascimento
et al. (2018) also performed the classification of CLL, FL and MCL
groups, extracting morphological and non-morphological features of
the cell nuclei. The authors applied several selection and classification
techniques. The best accuracy obtained was 100%. Ribeiro et al. (2018)
proposed the combination of fractal, curvelet and, Haralick features
to classify the NHL groups. The authors tested several classifiers such
as decision tree, random forest, SVM, Naive Bayes and K*, and it was
possible to obtain an accuracy rate of 86.14%.
Also, Brancati et al. (2019), Janowczyk and Madabhushi (2016),
Tambe et al. (2019), Zhang et al. (2021) and Bai et al. (2019) used deep
learning to classify NHL groups. The authors used convolutional neural
networks (CNN) and hand-crafted features to extract and classify image
patterns. Janowczyk and Madabhushi (2016) used an Alexnet model
with classification errors most commonly on low-quality images. Tambe
et al. (2019) performed the classification through an Inception V3
network with hyperparameters tuning. Brancati et al. (2019) used an
autoencoder with symmetric architecture and skip connections. This
approach reached higher accuracy than U-net and ResNet. In contrast,
the ResNet50 model was used by Zhang et al. (2021) with image
transformations for obtaining sufficient data for training, validation
and test of the networks. Bai et al. (2019) used a modified version of
GoogLeNet with statistical, texture and color features. Among all these
studies, it was possible to obtain accuracy rates between 96.58% and
99.33%.
Battula and Sharma (2018) presented a review of published works
of lymphoma histological images classification. This study highlight the
feature extraction by wavelet and Fourier transforms, Gabor filter and
gray level co-occurrence matrix. This matrix was also used by Taino
et al. (2020) along with percolation, lacunarity and fractal features.
The proposed methodology used a genetic algorithm for selecting the
best combination of features, selection algorithms, and classifiers. The
evolutionary algorithm allowed the selection of 73 features through
the Relief method with the k-nearest neighbors, reaching an area
under the curve of 0.947. Jiang et al. (2018) proposed a hierarchical
approach with images divided into 130 patches, with no overlapping.
The feature extraction was made by sparse autoencoder and texture
representations. The classification reached a 97.96% of accuracy with
two steps: the first one separated FL from the other lymphomas samples
G.B. Rozendo et al.
through an extreme learning machine, and the second one divided the
samples in CLL and MCL by using SVM algorithm.
Fractal features were also used by recent studies of lymphoma
images. Martins et al. (2019) applied these features with lacunarity,
analyzing different color models. The classification was performed by
Hermite polynomial with an accuracy of 97.60%. Candelero et al.
(2020) also performed a lymphoma images classification with fractal
and CNN features, but also with texture representations. The best
CNN features were selected by the ReliefF and evolutionary methods.
The proposed methodology reached a maximum accuracy of 82.01%
with K* classifier and the genetic algorithm. In comparison with other
analyzed images, it was possible to notice that these features better
represent breast cancer and colorectal cancer images. Roberto et al.
(2021) classified histological images with CNN and fractal features. To
do so, 300 fractal features were scaled and offered as input for the CNN.
Besides, another CNN was used with the original images as inputs. The
class probabilities of both networks were used for the definition of the
images classes. The classification obtained an accuracy of 95.55%, with
future possible parameters optimizations.
Even with the advances previously presented to support the devel-
opment of CAD systems dedicated to the context of NHL images, the
methods were defined with techniques based on Fourier and wavelet
transforms, morphological and non-morphological models, fractal anal-
ysis and deep learning strategies, for instance. However, the texture
irregularities and measure of the disorder of the histological structures
have not been verified through SampEn signatures and signature met-
rics, or even considering the descriptor of the behavior with multiple
segmentation and color normalization techniques. These insights and
those previously mentioned (Section 2.1) have been the main moti-
vations for this study. This type of investigation can provide a full
understanding of solutions to distinguish NHL groups, expanding the
quantitative capacity of SampEn and providing valuable information
for improving CAD systems, such as: association of SampEn signatures
and different classifiers for evaluating H&E images; evaluation of the
discriminative ability of SampEn signatures on NHL images, after apply-
ing multiple segmentation techniques and color normalization models;
evaluations based on signature metrics, enhancing the quantitative
capacity of the SampEn; indications of the best features, with tests
under different conditions.
3. Methods
The proposed method was divided into four main phases, as illus-
trated in Fig. 1. In order to investigate the performance of this proposal,
the first phase (Datasets: NHL images, Section 3.1) defines the three
histological datasets, with the original NHL images and the achieved
results with multiple segmentation and color normalization techniques.
The second phase (Feature extraction: sample entropy, Section 3.2)
performs the extraction of SampEn values, using distinct associations
of values for the parameters 𝑚 and 𝑟. From the SampEn values, the 𝑚
signatures curves with 𝑟 points were constructed. Finally, the attributes
area, skewness, maximum entropy point and area ratio were obtained
from each SampEn signature, representing the signature metrics. The
third phase (Classification, Section 3.3) performs three tests: classi-
fication provided by each SampEn value; a classification based on
SampEn signatures; and, classification using signature metrics from
SampEn. The feature vectors were classified using different types of al-
gorithms. The goal was to identify the combination (dataset, attributes
and classification approach) able to provide the best performance to
distinguish the NHL images, with the least number of features. The
best combination, in terms of accuracy, was observed under different
conditions, defining the fourth phase (Evaluation, Section 3.4). Each
phase is described in detail in the following sections.
The proposed method was implemented using the MATLAB® R2017a
and Weka 3.8.1 (Garner et al., 1995) packages. We have used the
default values suggested by the tools. The experiments were performed
on a computer with an Intel® CoreTM i5-3230M, 2.60 GHz, 3rd.
generation, 8 GB of RAM and Windows operating system with 64-bit
architecture.
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Expert Systems With Applications 202 (2022) 117238
3.1. Datasets: NHL images
The proposed method was evaluated using NHL images from a
public dataset, here identified as original H&E histological images,
provided from studies carried out by researchers at the National Cancer
Institute and National Institute on Aging, both located in the United
States (National Cancer Institute, 2022; National Institute on Aging,
2022; Shamir, Orlov, Eckley, Macura, & Goldberg, 2008). This is the
main dataset explored by important techniques available in the liter-
ature (Bai et al., 2019; Brancati et al., 2019; Codella et al., 2016; do
Nascimento et al., 2018; Janowczyk & Madabhushi, 2016; Meng et al.,
2010; Nascimento et al., 2015; Orlov et al., 2010; Ribeiro et al., 2018;
Roberto et al., 2021, 2017; Song et al., 2012; Taino et al., 2020; Tambe
et al., 2019). The dataset was defined from 30 histological slides of
lymph nodes stained with H&E, with ten cases for each group: FL;
CLL; and, MCL. Based on the characteristics of each group, to better
represent clinical practice, instead of the strictly controlled laboratory
environment, the slides were obtained with significant variations in
sectioning and staining, which can represent challenges for image
processing methods.
For this dataset, the microscopic images were digitally obtained
using a light microscope (Zeiss Axioscope) with a 20× objective and
a color digital camera (Axio Cam MR5) attached. Regions of inter-
est for each slide were selected by specialists, digitally photographed
and recorded without compression, RGB color model, resolution of
1388 × 1040 pixels. In total, 375 images were generated, contain-
ing 113, 140, and 122 cases of CLL, FL, and MCL, respectively. In
Fig. 2, examples of images corresponding to each of the groups are
provided. It is important to highlight that the RGB color model was
also used to represent the results obtained in the segmentation and
normalization methods. Then, for the feature extraction step, each RGB
image was converted into a corresponding grayscale image, with an 8-
bit quantization rate, in order to allow the application of the SampEn
technique.
3.1.1. Investigation considering segmentation techniques
In addition to the original NHL images, the proposal was tested
on the results provided by segmentation techniques. For this pur-
pose, the NHL images were segmented both by a specialist (gold
standard) (Tosta, Faria, et al., 2017) and using methods available in the
literature, which included the approaches explored by De Oliveira et al.
(2013), Vahadane and Sethi (2013), Wienert et al. (2012) and, Tosta,
Faria, et al. (2017). In addition to these previous works, we have also
applied recent approaches to perform the segmentation of the NHL
images, such as Tang et al. (2018), Tokime et al. (2018) and, Silva
et al. (2019). The methods were applied to define regions of interest
and investigate how these can influence the classification process of
NHL groups. We chose these methods because they were specifically de-
signed and/or tested to study nuclear structures in histological images
stained with H&E.
The methods based on (De Oliveira et al., 2013; Silva et al., 2019;
Tang et al., 2018; Tokime et al., 2018; Tosta, Faria, et al., 2017;
Vahadane & Sethi, 2013; Wienert et al., 2012) were evaluated with
metrics commonly explored by studies related to the segmentation pro-
cess of CLL, FL, and MCL groups: accuracy, sensitivity, and specificity.
These metrics were defined considering the concepts of true positive,
true negative, false positive, and false negative (Oger, Belhomme, &
Gurcan, 2012; Tosta, Neves, & do Nascimento, 2017). Thus, the accu-
racy value defines the percentage of correctly segmented pixels (nuclear
structures and background) when compared to manual segmentation
(specialist or gold standard, Tosta, Neves, & do Nascimento, 2017).
Sensitivity indicates how many pixels from manual segmentation of
nuclear structures were also identified by the computational approach.
Specificity indicates how many pixels defined as background by manual
segmentation were also identified by the automatic technique (Oger
et al., 2012; Tosta, Neves, & do Nascimento, 2017).
G.B. Rozendo et al.
Fig. 1. Overview of the proposed method.
Fig. 2. Examples of the histological images from the lymphomas groups: (a) CLL; (b) FL; and, (c) MCL.
The first method applied was the one proposed by Wienert et al.
(2012) which is a contour-based approach to cell segmentation. The
approach uses minimal a priori information, identifying contours re-
gardless of shape.
The method proposed by De Oliveira et al. (2013) is a three-step
segmentation process. In the first step, a contrast elongation technique
is applied on the R channel of the RGB color model. In the second
step, global thresholding based on minimum cross-entropy is applied,
and finally, in the third step, false positives are removed using the
connected components method.
The method proposed by Vahadane and Sethi (2013) is an improve-
ment on the seeded watershed segmentation method. First, foreground
seeds are generated through rapid radial symmetry transformation.
After that, the Otsu thresholding method is applied in the histograms
and background markers are calculated. Finally, false true detection
and removal are performed using Boolean operators.
The method proposed by Tosta, Faria, et al. (2017) consists of
an unsupervised method of segmentation of nuclear components of
neoplastic cells that combines histogram equalization, Gaussian filter,
fuzzy 3-partition entropy, genetic algorithm, morphological techniques,
and method of vouchers emphasis.
A strategy based on CNN was also applied in our investigation,
considering the study performed by Tokime et al. (2018). These authors
used the architecture described by Ronneberger, Fischer, and Brox
(2015) in order to define nuclei segmentation in H&E images. This
strategy uses a CNN architecture called UNet that performs semantic
segmentation. The UNet is an encoder–decoder network wherein the
encoder is a series of convolutional layers that learn to extract low-
resolution features, while the decoder allows precise localization of
high-resolution features.
Another CNN applied to provide segmented images in our study was
based on an approach presented by Tang et al. (2018). The authors
6
Expert Systems With Applications 202 (2022) 117238
explored the architecture described by Badrinarayanan, Kendall, and
Cipolla (2017) for evaluating the histological images. The method con-
sists of a CNN architecture that performs semantic pixel segmentation.
The CNN is composed of an encoder network, a decoder network, and
a pixel classification layer. The function of the decoder network is to
perform a conversion of the low-resolution feature maps, generated by
the encoder network, to high-resolution feature maps.
Finally, we used the Expectation–Maximization method (EM) with
the Gaussian Mixture Model (GMM), the EM-GMM model, as a strat-
egy to define the nuclei segmentation in H&E images (Silva et al.,
2019). The EM-GMM model was based on a proposal described by Ir-
shad, Veillard, Roux, and Racoceanu (2013), Ragothaman, Narasimhan,
Basavaraj, and Dewar (2016). Thus, the GMM creates probability den-
sity functions (PDF) to estimate the probability that each pixel belongs
to a class. The parameters of these PDF functions are estimated using
the EM method, which consists of steps E and M. Step E computes
the logarithmic probability of the entire dataset. Step M finds the
parameters by maximizing the logarithmic probability. These steps are
repeated until they converge, and the mean and covariance matrices
obtained in the convergence are chosen as the final parameters in the
model.
3.1.2. Investigation considering color normalization techniques
Another application context of the model aimed to define the dis-
criminative power of SampEn signatures on histological images after
applying color normalization techniques. Color normalization methods
are developed as preprocessing strategies to assist in correcting color
variations due to the staining process.
It is important to highlight that the H&E staining is useful for pattern
recognition and classification because the stains allow to highlight
cell structures of interest. Hematoxylin stains acid structures in blue–
purple shades, such as nuclei. Eosin stains basic pH components in pink
G.B. Rozendo et al. Expert Systems With Applications 202 (2022) 117238

shades, such as cytoplasm areas. The images obtained from this process Besides, histological image pixels represent a stain mixture, which
represent these conditions through colors, or different intensities when justifies the use of the sparsity approach. This estimation was made
observed in grayscale. Thus, the architecture of tissue components by a dictionary learning reaching a global optimum which allowed the
and cellular morphological aspects are highlighted, which are essential stain density map estimation through a robust sparse coding. Before
for tissue texture extraction and cancer diagnosis (Tosta, de Faria, these estimations, the RGB values of both source and target images
Neves, & do Nascimento, 2019; Tosta, Neves, & do Nascimento, 2017). were converted to the optical density space and after those, the nor-
However, the existing variations can also be caused by the non-uniform malized image was defined as performed by Macenko et al. (2009).
distribution of the H&E stains or by bad lighting conditions during the The RGB image was then obtained by application of the inverse of the
acquisition process (Feng et al., 2018; Jothi & Rajam, 2017). This condi- Beer–Lambert law.
tion can influence the interpretation and decision-making of specialists
and computer systems during the process of diagnosing a disease under 3.2. Feature extraction: sample entropy
investigation. For this reason, the color normalization techniques of
H&E images have been increasingly investigated to compose a new SampEn calculations were performed by considering that each im-
phase in the process of developing diagnostic support systems. The age is a matrix 𝑈𝑖𝑗 , and each position within the image was related to
color normalization consists of standardizing the colors of all images a grayscale value (Richman & Moorman, 2000; Silva et al., 2016; Yeh
that are part of a dataset (Jothi & Rajam, 2017). In this context, the et al., 2011). Sub-matrices (windows) 𝑥𝑚 𝑖,𝑗 sized 𝑚 × 𝑚, overlapped the
applied techniques were the models described by Khan et al. (2014), image sized 𝑁𝑈 = max(𝑖) × max(𝑗) with the goal of attaining samples of
Macenko et al. (2009), Reinhard et al. (2001) and, Vahadane et al. possible patterns contained in 𝑈𝑖𝑗 , are given by:
(2016), using the results presented by Tosta, de Faria, Servato, et al.
(2019). 𝑥𝑚
𝑖,𝑗 = {𝑢(𝑖, 𝑗), 𝑢(𝑖, 𝑗 + 1), … , 𝑢(𝑖 + 1, 𝑗), … , 𝑢(𝑖 + 𝑚 − 1, 𝑗 + 𝑚 − 1)}, (1)
The method proposed by Reinhard et al. (2001) is one of the most
applied in the literature. First, the images are converted from the wherein 1 ≤ 𝑖 ≤ max(𝑖)−𝑚+1 and 1 ≤ 𝑗 ≤ max(𝑗)−𝑚+1. To verify existing
RGB space to the 𝑙𝛼𝛽 space due to its minimum correlation among its similarities, each window 𝑥𝑚 𝑖,𝑗 was fixed in position (𝑖, 𝑗) and compared
channels in natural images. This condition allows that color alterations to windows 𝑦𝑚 𝑖′ ,𝑗 ′
, which were positioned throughout the image.
using this model do not lead to artifacts in the resulting image. The The similarities between two patterns were calculated from a dis-
𝑚 between two windows 𝑥𝑚 and 𝑦𝑚 . The distance was defined
tance 𝑑𝑥,𝑦
metrics of mean and standard deviation are calculated for each 𝑙𝛼𝛽 𝑖,𝑗 𝑖′ ,𝑗 ′
channel for the target and source images. The new 𝑙𝛼𝛽 channels of the as the maximum difference among the pixel intensity of the windows:
source image are then computed by all these metrics. In the end, the ′ ′
𝑖,𝑗 , 𝑦𝑖′ ,𝑗 ′ ] = max |𝑢(𝑖 + 𝑘, 𝑗 + 𝑙) − 𝑢(𝑖 + 𝑘, 𝑗 + 𝑙)|,
𝑚
𝑑𝑥,𝑦 = 𝑑[𝑥𝑚 𝑚
(2)
𝑙𝛼𝛽 channels are converted to the RGB model. Although it was proposed
for natural images, the normalization of Reinhard et al. (2001) is wherein 𝑘 > 0, 𝑙 < 𝑚 − 1 and 𝑖, 𝑗 ≠ 𝑖′ , 𝑗 ′ , not including self-comparisons.
commonly used by studies of histological images processing (Tosta, The probability of a window 𝑦𝑚 𝑖′ ,𝑗 ′
being at a maximum distance of 𝑟
de Faria, Neves, & do Nascimento, 2019). from a fixed window 𝑥𝑚 𝑖,𝑗 was defined by determining the average count
Another approach applied was following the method presented of all windows 𝑦𝑚𝑖′ ,𝑗 ′
that were close to 𝑥𝑚 𝑖,𝑗 , according to Eq. (3).
by Macenko et al. (2009). In this approach, the RGB colors in images
are converted to the optical density space. This space allows linear 1 ∑ max(𝑗)−𝑚
max(𝑖)−𝑚 ∑
𝐶 𝑚 (𝑥𝑚
𝑖,𝑗 ) = 𝜃(𝑑[𝑥𝑚 𝑚
𝑖,𝑗 , 𝑦𝑖′ ,𝑗 ′ ] − 𝑟), (3)
combination of the stains to be also linearly represented. In this repre- 𝐷−1 𝑖′ =1 𝑗 ′ =1
sentation, each RGB channel is converted to its corresponding optical
densities by calculating the logarithm of its normalized value. This with (𝑖, 𝑗) ≠ (𝑖′ , 𝑗 ′ ), 𝐷 = (max(𝑖) − 𝑚) × (max(𝑗) − 𝑚) and 𝜃(𝑑[𝑥𝑚 𝑚
𝑖,𝑗 , 𝑦𝑖′ ,𝑗 ′ ] − 𝑟)
processing allows the estimation of a stain color appearance matrix and the Heaviside function defined by:
{
a stain density map. The stain color appearance matrix specifies the rep- 1, if 𝑑𝑥𝑦 𝑚 − 𝑟 ≤ 0,
𝑚
resentation of each stain by the RGB channels and the stain density map 𝜃(𝑑𝑥𝑦 − 𝑟) = (4)
0, if 𝑑𝑥𝑦 𝑚 − 𝑟 > 0,
quantifies the saturation of each stain in each pixel (Tosta, de Faria,
Neves, & do Nascimento, 2019). To estimate these components, optical wherein 𝑟 is a tolerance parameter that defines whether two patterns
densities were used to estimate the color appearance matrix by the are considered similar. Two patterns are considered similar if each
singular value decomposition method. The density map could be then corresponding pixel within the patterns is distant at most 𝑟 from
calculated using the optical densities and the matrix coefficients. These each other (Silva et al., 2016). Thus, the value of 𝑟 is determined by
estimates were performed for the target and source images, obtaining evaluating the standard deviation of the analyzed image, given by:
a normalized image through combination between the target image
matrix and the source image map. 𝑟 = 𝛽 × 𝜎(𝑈 ), (5)
The stain color appearance matrix was also used by Khan et al. in which 𝛽 is the real positive constant and 𝜎 is the standard deviation
(2014). This technique estimates the color appearance matrix by a of image 𝑈 .
classification of pixels into hematoxylin, eosin, and background using SampEn was calculated using the negative of natural logarithm of
an oct-tree quantization, RGB values, and the relevance vector machine the ratio between similarity coefficients of all analyzed patterns of size
classifier. This processing was performed for both target and source (𝑚 + 1) × (𝑚 + 1) with similarity coefficients of patterns 𝑚 × 𝑚:
images, which were deconvolved for further nonlinear mapping be-
tween them. To do so, the metrics of mean, 5th percentile, and 95th 𝛷𝑚+1
𝑆𝑎𝑚𝑝𝐸𝑛(𝑚, 𝑟, 𝑁𝑈 ) = − ln , (6)
percentile were calculated for each channel of both images and used for 𝛷𝑚
mapping these metrics of the source image to the target image through wherein 𝛷𝑚+1 and 𝛷𝑚 are the similarity coefficients defined respec-
the B-spline method. The normalized image was obtained by the target tively by:
color appearance matrix combined with the stain channels of the source
1 ∑
max(𝑖)−(𝑚+1) ∑
max(𝑗)−(𝑚+1)
image. 𝛷𝑚+1 = 𝐶 𝑚+1 (𝑥𝑚+1 (7)
𝐷 𝑖,𝑗 ),
Vahadane et al. (2016) presented an estimation of the stain color 𝑖=1 𝑗=1
appearance matrix by a sparse non-negative matrix factorization. This and
method was chosen since the matrix coefficients cannot have negative
values, which would be biologically incorrect because this condition 1 ∑ max(𝑗)−𝑚
max(𝑖)−𝑚 ∑
𝛷𝑚 = 𝐶 𝑚 (𝑥𝑚
𝑖,𝑗 ). (8)
represents a light-emitting that is not possible in the tissue staining. 𝐷 𝑖=1 𝑗=1

7
G.B. Rozendo et al.
Table 1
Average standard deviation values obtained from different combinations of 𝑣 sub-images sized 𝑛 × 𝑛.
𝑛 𝑣
20 40 60 80 100
30 0.0680 0.0489 0.0421 0.0362 0.0335
40 0.0597 0.0423 0.0341 0.0295 0.0286
In this paper, SampEn was applied in order to use sub-images to
enable the quantification of images. This step is necessary to determine
SampEn values, since the quantity of comparisons among pixels was on
the order of 2×(max(𝑖)−𝑚+1)2 ×(max(𝑗)−𝑚+1)2 (Yeh et al., 2011). The
selected procedure considered 𝑣 sub-images of size 𝑛 × 𝑛, which were
randomly selected throughout different positions of the input image
𝑈 , following a uniform distribution model. After calculating values for
each sub-image, a SampEn value for the whole image was obtained
using the simple arithmetic average of the entropy values of all chosen
sub-images (𝑣).
𝑆𝑎𝑚𝑝𝐸𝑛1 (𝑚, 𝑟, 𝑛) + ⋯ + 𝑆𝑎𝑚𝑝𝐸𝑛𝑘 (𝑚, 𝑟, 𝑛)
𝑆𝑎𝑚𝑝𝐸𝑛𝑈 (𝑚, 𝑟, 𝑁𝑖𝑚𝑔 ) = , (9)
𝑣
in which 𝑆𝑎𝑚𝑝𝐸𝑛𝑈 represents the entropy of the image and 𝑆𝑎𝑚𝑝𝐸𝑛1 ,
⋯, 𝑆𝑎𝑚𝑝𝐸𝑛𝑘 are entropies of the sub-images 1 to 𝑘, respectively.
The values of 𝑛 and 𝑣 used for quantifying the NHL images were
defined considering a test that was based on standard deviation val-
ues (Yeh et al., 2011). The test involved calculating SampEn repeatedly
for the images included within the lymphoma dataset, and considering
different values for parameters 𝑛 and 𝑣. Also, the standard deviation
of SampEn values were calculated. The acceptable values of 𝑛 and 𝑣
were calculated from tests performed on an image subset from National
Cancer Institute (2022) and National Institute on Aging (2022). This
subset was comprised of approximately 10% of the available images
of each group. The images were randomly chosen. Each combination
(𝑛, 𝑣) was tested 10 times per image, without window overlapping,
which made it possible to obtain the average value of SampEn and the
standard deviation of the chosen image. This process was applied to all
images within the subset. Thus, the combination (𝑛, 𝑣) that resulted in
standard deviation value below 0.02 was selected for quantifying all
images of the lymphoma dataset. Sufficiently small standard deviation
value indicates that SampEn value was acceptable for quantifying
the image. This type of analysis and standard deviation thresholding
procedure were previously defined by Yeh et al. (2011). Examples of
combinations used are presented on Table 1, which includes average
standard deviation values for each pair (𝑛, 𝑣). The best combination
with the imposed requirement was obtained with 𝑛 = 40 and 𝑣 = 180,
and provided an average standard deviation of 0.0198. These values
were selected for the study of lymphoma images.
3.2.1. SampEn signatures
A texture signature is useful for quantifying the spatial arrange-
ment and intensities of pixels in a digital image. In our proposal,
this was made possible by computing SampEn values from multiple
observations, which involved quantifications using different combina-
tions of values for the parameters 𝑚 and 𝑟. The parameter 𝑚 was
employed to determine pixel window sizes in order to assess patterns
within an image. The parameter 𝑚 was derived from the set N∗ and
represented the observation dimension. It is important to note that
the greater the 𝑚 value, the greater the possibility of no pattern
matches (Humeau-Heurtier, 2019). Thus, the lack of similarities be-
tween windows produces division by zero in Eq. (6). In this paper,
the quantifications were performed using 𝑚 values ranging from 1 to
4, at intervals of one unit, in order to avoid this problem. Also, the
parameter 𝑟 was applied to indicate the tolerance level so that two
windows sized 𝑚 are defined as similar. Similarity was defined as the
𝑚 between two windows (𝑥𝑚 and 𝑦𝑚 ) according to Eq. (2).
distance 𝑑𝑥,𝑦
𝑖,𝑗 𝑖′ ,𝑗 ′
The parameter 𝑟 was a real number greater than zero. Therefore, the
signatures were obtained for values of 𝑟 between 0.06 and 0.4, at
8
Expert Systems With Applications 202 (2022) 117238
120 140 160 180
0.0282 0.0272 0.0252 0.0243
0.0253 0.0222 0.0222 0.0198
increments of 0.02. The chosen values (𝑚 and 𝑟) were widely discussed
within important works available in the literature, such as by da Silva
et al. (2014), Silva et al. (2016), and Richman and Moorman (2000).
A signature of 𝑚 = 1 and 𝑟 = 0.06 provided the quantification of
minor patterns contained within an image, and indicated increases in
restrictive tolerance. On the other hand, a signature obtained using
𝑚 = 4 and 𝑟 = 0.04 facilitated the quantification of major patterns, and
indicated a decreasing restrictive tolerance. Quantifications obtained
using SampEn were organized as curves (texture signatures) in the
coordinate system SampEn (Y axis) × 𝑟 (X axis). Signatures were applied
to quantify texture complexity for different values of 𝑚, and provided
a more complete and accurate description of the studied images. For
each image, four texture signatures were obtained, one for each value
of 𝑚.
3.2.2. Signature metrics
Each SampEn signature was quantitatively evaluated from met-
rics (Căliman & Ivanovici, 2012), with the goal of quantifying the
behaviors of each NHL group as a function of multiple observations
of 𝑚. The applied metrics included area under curve (𝐴), skewness (𝑆),
maximum point of entropy (𝑃𝑚𝑎𝑥 ) and area ratio (𝑅).
The metric 𝐴 was defined as the existing area between a function,
𝑓 (𝑥), and axis 𝑥, on an interval [𝑎, 𝑏]. A value of 𝐴 was obtained
by calculating the integral of 𝑓 (𝑥) for range [𝑎, 𝑏]. However, since
SampEn is a discrete function, the value of 𝐴 was computed from a
trapezoid-based approach (Mathworks, 2022b):
𝑁𝑒
1∑
𝐴≈ (𝐸 − 𝐸𝑖 )[𝑓 (𝐸𝑖 ) + 𝑓 (𝐸𝑖+1 )], (10)
2 𝑖=1 𝑖+1
wherein (𝐸𝑖+1 − 𝐸𝑖 ) indicates the spacing between two consecutive
points, 𝑁𝑒 is the number of entropy values of the curve and 𝑓 (𝐸𝑖 ) is
the entropy value of point 𝐸𝑖 .
The metric 𝑆 indicated curve asymmetry relative to its average
value. If the values of the curve are more concentrated to the right
of the curve average, skewness is positive. When values of the curve
are more concentrated to the left, skewness is negative. Skewness was
calculated according to Eq. (11) (Mathworks, 2022a):
∑ 𝑁𝑒
1
𝑖=1
𝑁𝑒
(𝑓 (𝐸𝑖 ) − 𝑓 (𝐸))3
𝑆= √ , (11)
∑ 𝑁𝑒
( 𝑁1 𝑖=1
(𝑓 (𝐸 𝑖 ) − 𝑓 (𝐸))2 )3
𝑒
wherein 𝑓 (𝐸𝑖 ) is the 𝑖th entropy value, 𝑓 (𝐸) is the curve average and
𝑁𝑒 is the number of points in the curve.
The metric 𝑃𝑚𝑎𝑥 made it possible to acquire the highest degree of
entropy for each signature obtained for each function of 𝑚.
The metric 𝑅 was determined from the average value of each
analyzed signature. A 𝑅 value was calculated as the ratio between areas
on the left and right sides relative to the center of the curve, according
to the following Equation:
𝐴(𝑁𝑒 ∕2+1,𝑁𝑒 )
𝑅= , (12)
𝐴(1,𝑁𝑒 ∕2)
in which 𝑁𝑒 indicates the number of points in the curve.
G.B. Rozendo et al.
Fig. 3. Illustration of the feature vector structure considering the metrics 𝐴, 𝑆, 𝑃𝑚𝑎𝑥
and 𝑅 for each signature (𝑚 ranging from 1 to 4).
3.2.3. Compositions based on SampEn features
The SampEn values were obtained from the three datasets (original
histological images and those resulting from the multiple segmentation
and color normalization techniques). Thus, the values were used by
exploring different strategies. The first one considered all the SampEn
values, which were calculated for each combination of 𝑚 and 𝑟. This
strategy explored the features defined as SampEn signature. The result
is a set with 72 SampEn values, that were produced using 𝑚 values
that ranged from 1 to 4 and 𝑟 values that ranged from 0.06 to 0.40.
The second strategy aimed to identify the discriminative power of each
SampEn value (analyzed individually). This type of test is the most
frequently described in the literature (da Silva et al., 2014; Richman
& Moorman, 2000; Silva et al., 2016). Here, this type of test was
identified as classification provided by each SampEn value. Finally,
the signature metrics (𝐴, 𝑆, 𝑃𝑚𝑎𝑥 and 𝑅) were calculated for each
SampEn signature, totaling 16 attributes, as illustrated in Fig. 3. This
last strategy represents another important contribution of this work
considering the quantitative power of SampEn in the context of NHL
images.
3.3. Classification
The classifications were performed through different strategies, such
as support vector machine - SVM (function-based), K* (lazy learning)
and random forest (decision tree-based). Also, evolutionary approaches
were applied in order to define the most relevant features in each tested
combination, with a full understanding of the solutions to distinguish
the NHL groups. The chosen strategies were genetic algorithm (genetic
evolution-based), particle swarm optimization (particle swarm-based)
and binary gray wolf optimization (animal behavior-based), all using
the K-nearest neighbor (KNN) classifier. KNN is a commonly used
method for fitness function due to its simplicity, fast processing speed
and high classification performance (Emary, Zawbaa, & Hassanien,
2016; Hans & Kaur, 2022; Li, Xue, & Zhang, 2021; Maleki, Zeinali, &
Niaki, 2021; Too & Abdullah, 2021).
Each classification strategy was evaluated using the weka v3.8.1
package (Garner et al., 1995) and the corresponding algorithms were
SMO (Sequential Minimal Optimization), KStar, RandomForest, Genet-
icSearch, PSOSearch and WolfSearch. The performance metric used to
evaluate the tests was accuracy (𝐴𝑐𝑐), according to Eq. (13). Also,
since some of the NHL groups as well as the different types of images
explored have a small number of samples, we chose to apply 3-fold
cross-validation in all testing stages in order to avoid problems such as
overfitting.
𝑇𝑃 + 𝑇𝑁
𝐴𝑐𝑐 = , (13)
𝑁𝐶
wherein 𝑇 𝑃 is the number of cases classified as true positives, 𝑇 𝑁 is
the number of cases classified as true negatives and 𝑁𝐶 is the total
number of cases.
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Expert Systems With Applications 202 (2022) 117238
The SVM algorithm is a deterministic supervised classification
method, wherein each instance is perceived as a point in a space with
a dimension that is equal to the number of extracted features. This
technique allows researchers to define a hyperplane that provides an
optimal degree of point separation between distinct classes, which have
the greatest possible distance between the hyperplane and its nearest
points. There are different approaches for defining the hyperplane,
which are known as kernel functions (Platt, 1999). The used parameters
were 𝐶 = 1, 𝜖 = 10−12 and polynomial kernel.
K* associates a class to an instance within a test set from classes of
the training set using similarity. For each test instance, a comparison
between the whole training set is performed, while considering entropy
as a criterion. The class that provides the highest complexity is defined
as the class of the new instance (Cleary & Trigg, 1995). The value of
20% was used as the global bend parameter.
The random forest (RF) algorithm combines several decision trees in
order to perform class prediction (Breiman, 2001). A decision tree is a
statistical algorithm composed of nodes connected by edges wherein
internal nodes represent decisions derived from a rule and leaves
represent the possible classifications. A tree is built for each class of the
training set. Each instance of the set is evaluated in relation to the built
trees, using a process based on voting. The class that obtains the most
votes is specified as the class of the studied instance. The parameters
used were maximum depth equal to 0, size of each bag equal to 100%,
100 iterations and minimum number of instances in the leaves equal
to 1.
The genetic algorithm (GA) uses an optimal or sub-optimal solution,
which is sought using techniques inspired by the process of natural
selection in evolutionary biology (Pei, Goodman, Punch, & Ding, 1995).
Natural selection consists in the evolution of generations of individuals,
and is designed to eliminate weak individuals and retain the ones con-
sidered strong. In the algorithm, possible solutions contain subgroups
of features called chromosomes. For each generation, chromosomes are
subjected to the processes of crossover and mutation. The best chromo-
somes are passed forward the next generation. The KNN algorithm was
used to evaluate the classification power of each generation. This cycle
was repeated until the convergence toward an optimal solution was
achieved. The parameters used a crossover probability equal to 60%,
maximum number of generations equal to 20, mutation probability
equal to 3.3% and population size of 20.
Particle swarm optimization (PSO) is based on bird flocking and fish
schooling behaviors. The process provides means to search an optimal
solution via information exchange between individuals of a popula-
tion (Kennedy, 2011). In the algorithm, each solution is considered to
be a particle with characteristics that define its direction of movement.
These characteristics include its current position, objective function
and speed vector. The objective function is applied to determine the
particle with the highest discriminative power in the current iteration,
as evaluated by the KNN classifier. Through information exchange,
the other particles follow the movements of the best particles. This
behavior is repeated until the optimal solution is found. The used
parameters were individual weights equal to 0.34, inertia equal to
0.33, mutation probability of 1%, population size of 20 individuals, 20
iterations and social weight of 0.33.
Binary gray wolf optimization (bGWO) is an optimization method
that is based on the hunting strategy of gray wolves (Emary et al.,
2016). The hunting tactic of a pack consists of following, getting
closer, chasing and surrounding the prey until it stops moving. In the
mathematical model, the wolves are candidate solutions of subgroup
features. Initially, the positions of wolves were randomly set. The best
candidates were selected according to fitness functions, which were
evaluated according to the classification power of the solutions within
the current iteration, which was tested using the KNN classifier. For
each iteration, positions of wolves were updated until the best solution
was found (Emary et al., 2016; Mirjalili, Mirjalili, & Lewis, 2014). The
parameters used for the tests were a population of 20 individuals, 20
iterations, and absorption coefficient equal to 0.001, escape probability
of 80%, chaotic coefficient equal to 4, mutation probability of 1% and
minimum beta of 0.33.
G.B. Rozendo et al.
Fig. 4. Examples of segmented images obtained from a CLL sample (a), (Shamir et al., 2008). The images represent the segmentation results performed by a specialist (b) and
after applying the techniques described by De Oliveira et al. (2013) (c), Vahadane and Sethi (2013) (d), Wienert et al. (2012) (e), Tosta, Faria, et al. (2017) (f), Tokime et al.
(2018) (g), Tang et al. (2018) (h), and Silva et al. (2019) (i).
3.4. Evaluation
The discriminative capacity of the best combination was observed in
different situations in order to verify the contributions and limitations.
Details are presented in the next subsections.
3.4.1. Applying different levels of noise
The best combination of SampEn features and classifier was ana-
lyzed by intentionally applying different types of noise, as described
by Feng et al. (2018) and Zhu and Wu (2004). These tests are important
for determining the capacity of the model to classify CLL, FL and
MCL groups under variable conditions. Thus, each test was evaluated
considering the accuracy metric and noise levels of 20%, 40% and,
80%, as described by Martins et al. (2017), Xiao, He, Jiang, and Liu
(2010) and, Zhu and Wu (2004).
Considering the model presented by Zhu and Wu (2004), the best
combination of features was analyzed using the training and test sets
(𝐶𝑡𝑟𝑎𝑖𝑛 and 𝐶𝑡𝑒𝑠𝑡 ) that were defined using the 𝑘-fold cross-validation
technique, with a value of 𝑘 = 3. The clean training and test data
were used to compose their corresponding sets with noise (dirty). The
𝐷𝑡𝑟𝑎𝑖𝑛 and 𝐷𝑡𝑒𝑠𝑡 files were generated by applying different levels of noise
over the 𝐶𝑡𝑟𝑎𝑖𝑛 and 𝐶𝑡𝑒𝑠𝑡 data. Features affected by noise were randomly
selected. Therefore, a new noisy training set 𝐷𝑡𝑟𝑎𝑖𝑛 was obtained from
𝐶𝑡𝑟𝑎𝑖𝑛 and applied to distinguish instances of the test set 𝐶𝑡𝑒𝑠𝑡 : 𝐷𝑡𝑟𝑎𝑖𝑛 ×
𝐶𝑡𝑒𝑠𝑡 . The same percentage of noise was included in the test set to
generate the noisy test 𝐷𝑡𝑒𝑠𝑡 . After this procedure, the comparisons also
involved 𝐶𝑡𝑟𝑎𝑖𝑛 ×𝐷𝑡𝑒𝑠𝑡 and 𝐷𝑡𝑟𝑎𝑖𝑛 ×𝐷𝑡𝑒𝑠𝑡 , which represented clean training
feature sets against noisy test feature sets, as well as noisy training
feature sets against noisy test feature sets.
Finally, we also tested the best combination of SampEn features and
classifiers under image noise conditions. We applied well-known noises
to the field of histopathological imaging: random pixel corruption
(RPC) and white Gaussian noise (WGN) (Feng et al., 2018). The random
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Expert Systems With Applications 202 (2022) 117238
pixel corruption was applied by randomly selecting pixels from the
image and replacing them with values following a uniform distribution
over zero and the largest image pixel value. The white Gaussian noise
was also added by randomly selecting pixels but replacing them with
its values plus a value following the standard normal distribution with
zero of mean and 1 of standard deviation. Each test was evaluated after
adding noise in relation to the total pixels of the image.
3.4.2. Indications of comparisons with previous works and distinct texture
descriptors
All the tests explored here are important to indicate the best combi-
nation of techniques for the classification of NHL groups. In addition,
we compared our best result to the performances provided by other
techniques that explored the NHL dataset. The techniques were pre-
sented by Bai et al. (2019), Brancati et al. (2019), Candelero et al.
(2020), Codella et al. (2016), do Nascimento et al. (2018), Janowczyk
and Madabhushi (2016), Jiang et al. (2018), Martins et al. (2019),
Meng et al. (2010), Orlov et al. (2010), Ribeiro et al. (2018), Roberto
et al. (2021, 2017), Song et al. (2012), Taino et al. (2020), Tambe
et al. (2019), and Zhang et al. (2021), all exploring different combina-
tions from that applied in our proposal. In general, these approaches
were based on Fourier and wavelets transforms, Gabor filter, local
binary pattern, morphological features, fractal approaches, or convo-
lution kernels, as well as different classifiers (support vector machine,
random forest, DECORATE, and Polynomial), or deep learning-based
methods. Finally, comparisons with different texture descriptors were
also performed, especially considering the techniques based on mul-
tidimensional and multiscale quantifications, such as fractal dimen-
sional (Ivanovici & Richard, 2011), lacunarity (Căliman & Ivanovici,
2012; Ivanovici, Richard, & Decean, 2009) and percolation (Roberto
et al., 2021, 2017). These methods were applied to analyze the different
NHL images (original, segmented and normalized).
G.B. Rozendo et al. Expert Systems With Applications 202 (2022) 117238

Fig. 5. Examples of images obtained after applying color normalization techniques to a CLL sample (Fig. 4(a)). The results were defined from the proposals of Reinhard et al.
(2001) (a), Macenko et al. (2009) (b), Khan et al. (2014) (c), and Vahadane et al. (2016) (d).

Fig. 6. Accuracy values: based on total pixels segmented correctly (nuclear structures and background) by computational methods when compared to manual segmentation
(specialist, Tosta, Faria, et al., 2017), considering CLL (a), FL (b) and MCL (c) groups.

4. Results according to the results presented by Tosta, de Faria, Servato, et al.

The best combination of techniques was determined after analyzing
the original NHL images and the achieved results with multiple segmen-
tation and color normalization techniques. Thus, the results were based
on images segmented by a specialist (gold standard) (Tosta, Faria,
et al., 2017) and computational techniques (De Oliveira et al., 2013;
Silva et al., 2019; Tang et al., 2018; Tokime et al., 2018; Vahadane
& Sethi, 2013; Wienert et al., 2012). Fig. 4 shows the segmented
images obtained from a sample of the CLL group (Fig. 4(a)). Also,
color normalization techniques were applied to assist in correcting
color variations due to the staining process (Jothi & Rajam, 2017). The
explored models were the proposed by Khan et al. (2014), Macenko
et al. (2009), Reinhard et al. (2001), and Vahadane et al. (2016),
(2019). Fig. 5 shows normalized images obtained from a histological
sample of the CLL group (Fig. 4(a)).
It is important to highlight that segmentation results can be eval-
uated through different metrics. Thus, in relation to the segmentation
quality achieved with each technique in NHL images, an overview is
illustrated in Figs. 6–8, considering box plots of accuracy, sensitivity,
and specificity, respectively, for all kinds of investigated NHL types.
These metrics were described in Section 3.1.1. Regarding the normal-
ization process for NHL images, the quality achieved in each technique
was presented in detail by Tosta, de Faria, Servato, et al. (2019).
Considering the segmentation results, Fig. 6 illustrates that the
best median accuracies were reached by the techniques of Tokime
et al. (2018) and Tang et al. (2018) in the images of all NHL groups,

11
G.B. Rozendo et al. Expert Systems With Applications 202 (2022) 117238

Table 2
Mean values of SampEn for the groups of lymphomas CLL, FL, and MCL.
𝑚=1 𝑚=2 𝑚=3 𝑚=4
𝑟 = 0.06 ⋯ 𝑟 = 0.12 𝑟 = 0.14 ⋯ 𝑟 = 0.20 𝑟 = 0.22 ⋯ 𝑟 = 0.32 ⋯ 𝑟 = 0.28 ⋯ 𝑟 = 0.32 ⋯ 𝑟 = 0.40
CLL 5.9781 ⋯ 4.3801 3.9192 ⋯ 3.3062 3.0523 ⋯ 2.2824 ⋯ 2.4784 ⋯ 2.2972 ⋯ 2.0000
FL 5.1097 ⋯ 3.2908 2.9588 ⋯ 2.4218 2.2832 ⋯ 1.7130 ⋯ 1.9555 ⋯ 1.7968 ⋯ 1.5294
MCL 5.8143 ⋯ 4.1413 3.7120 ⋯ 3.0904 2.8915 ⋯ 2.1704 ⋯ 2.3483 ⋯ 2.1922 ⋯ 1.9043

Fig. 7. Sensitivity values: based on total pixels defined as nuclear structures by computational methods when compared to manual segmentation (specialist, Tosta, Faria, et al.,
2017), considering the CLL (a), FL (b) and MCL (c) images.

with values beyond 80%. This demonstrates that, among the evaluated
techniques, those based on CNN were capable of identifying in a higher
proportion nuclear and background regions as such, in comparison with
the specialist segmentation. These techniques also reached the highest
minimum values in all NHL classes, with the exception of some outliers
in the MCL class (Fig. 6(c)).
The sensitivity results illustrated in Fig. 7 show that the tech-
niques of De Oliveira et al. (2013) and Silva et al. (2019) reached
the highest median values in all classes, with results higher than
60%. This condition demonstrates that nuclear regions segmented by
the specialist were also identified by these techniques. In contrast,
these methods obtained the lowest median values of specificity in all
NHL groups, as illustrated by Fig. 8. This condition indicates that
these segmentation approaches did not identify the background like
the specialist. Through the joint analysis of sensitivity and specificity
results, it is possible to note that these techniques obtained a high
true positive rate (nuclei) and a low true negative rate (background),
which characterizes an over-segmentation, with nuclear areas bigger
than the gold standard. The highest specificity values were obtained
by the techniques of Tokime et al. (2018) and Tang et al. (2018) in all
classes. However, the sensitivity results obtained by these techniques
indicate an under-segmentation, with low true positive rates (nuclei)
and high true negative rates (background). The segmentation meth-
ods of Wienert et al. (2012) and Tosta, Faria, et al. (2017) reached
intermediate results in all NHL classes and all metrics.
4.1. SampEn values applied to distinguish NHL groups
The proposed method was applied to quantify histological NHL sam-
ples (original, segmented, and normalized) and the results consisted of
SampEn values, according to the (𝑚, 𝑟) parameters, for each image given
as input. For instance, Table 2 displays the averages of the SampEn
values that were calculated for the original images from the CLL, FL,
and MCL groups. Also, to illustrate the statistical significance of these
features, the Wilcoxon–Mann–Whitney test was applied to the SampEn
values that represent the different NHL groups. The significance level
was of 0.05. The results were mean 𝑝-values of 0.0036 (CLL versus
FL), 0.1458 (CLL versus MCL), and 0.0064 (FL versus MCL). These

12
G.B. Rozendo et al.
Fig. 8. Specificity values: based on total pixels defined as background by computational methods when compared to manual segmentation (specialist, Tosta, Faria, et al., 2017),
considering the CLL (a), FL (b) and MCL (c) images.
results are useful to show that the SampEn values present statistically
relevant differences for the NHL groups, mainly considering CLL × FL
and FL × MCL. Thus, the SampEn values can be analyzed using different
classifiers, according to the proposed methodology.
From the values obtained with SampEn, the signatures were con-
structed for different observations of 𝑚. For example, Fig. 9 displays
the SampEn signatures obtained from the images indicated in Fig. 2.
Notably, signatures of an image are different for each observation of
𝑚. Moreover, considering the observation 𝑚 = 1, the SampEn produced
values that were initially at the same tolerance level of 𝑟 = 0.06,
regardless of group (Fig. 9(a)). This did not occur for other observations
of 𝑚, in which SampEn did not identify similarities among the ana-
lyzed windows and, consequently, each class presented SampEn values
from distinct tolerance levels 𝑟. These characteristics are illustrated in
Figs. 9(b) (𝑚 = 2), 9(c) (𝑚 = 3) and 9(d) (𝑚 = 4). This behavior has
been properly discussed by Richman and Moorman (2000) and Azami,
Escudero, and Humeau-Heurtier (2017). Thus, the features obtained
can indicate that distinct patterns were quantified in the signatures of
each NHL class.
Tests were defined to show the superiority of texture signatures
for quantifying and classifying NHL groups, according to the strategies
presented in Section 3.2.3. The compared groups were CLL × FL, CLL
× MCL, FL × MCL and CLL × FL × MCL. Table 3 shows the best results
obtained from the SampEn signatures. The highest 𝐴𝑐𝑐 value for each
investigated situation is highlighted in bold. The second strategy was
applied to analyze the discriminative power of each SampEn value
(analyzed individually). The best results obtained with this strategy
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Expert Systems With Applications 202 (2022) 117238
are shown on Table 4, considering the different types of images and
NHL groups. The highest 𝐴𝑐𝑐 value for each investigated comparison is
highlighted in bold.
Considering the results shown on Tables 3 and 4, the highest 𝐴𝑐𝑐
rates were achieved from images segmented by a specialist, whether
involving the signature vectors or the classification provided by each
SampEn value, even when the images were segmented with CNN
models (Tang et al., 2018; Tokime et al., 2018). Also, taking as a main
goal the distinction among the three classes, CLL × FL × MCL, the best
result was 87.86% of 𝐴𝑐𝑐, exploring the strategy of SampEn signatures
with the bGWO method. This strategy indicates that the use of SampEn
signatures contributes to the process of classifying the NHL groups,
especially in comparison with the results provided through a strategy
commonly explored in the specialized literature on the topic (da Silva
et al., 2014; Richman & Moorman, 2000; Silva et al., 2016). For
example, when results via the classification of each SampEn value are
considered (Table 4), the highest 𝐴𝑐𝑐 value was of 72.25% with the
K*, GA, PSO, and bGWO methods. Finally, the classifications involving
comparisons of CLL × FL × MCL with original or normalized images
provided less expressive results than those previously highlighted.
4.2. Analysis of the NHL groups via metrics obtained from the SampEn
signatures
Another way to verify the discriminative power of SampEn signa-
tures was through metrics extracted from the curves. Thus, the metrics
were calculated from the SampEn signatures (Section 3.2.1) and the
G.B. Rozendo et al.
Fig. 9. Examples of SampEn signatures for the histological images shown in Fig. 2. The patterns were calculated with 𝑟 ranging from 0.06 to 0.4 for the observations 𝑚 = 1 (a),
𝑚 = 2 (b), 𝑚 = 3 (c), and 𝑚 = 4 (d).
results were provided as inputs for distinct classifiers (Section 3.3).
The performance provided via signature metrics from SampEn are
shown on Table 5, with indications in bold of the best rates (𝐴𝑐𝑐)
for each situation investigated. The classifications were performed
considering the original histological images and the corresponding
results obtained after applying the multiple segmentation and color
normalization methods.
From Table 5, it is possible to observe some highlights regarding
the use of metrics obtained from the SampEn signatures. For instance,
these results are relevant compared to those previously achieved, with
emphasis on the classifications using original histological images (non-
segmented and non-normalized). This is an important contribution
regarding how SampEn can be better explored for the process of quan-
tification and recognition of complex textures, such as NHL images. In
this combination, the 𝐴𝑐𝑐 rates were greater than 98% in all compar-
isons. The best performance occurred in the CLL × FL classification,
with an 𝐴𝑐𝑐 rate of 99.60%. Thus, the quantifications using the signa-
ture metrics from SampEn were able to distinguish different patterns
of NHL, even when different classifiers were applied. However, the
signature metrics from the segmented or normalized images indicated
classifications with less expressive performances. For these datasets,
the obtained results illustrate a possible limitation of this approach.
A possible explanation is that the patterns defined by the spatial and
textural arrangements (colors) were degraded from the segmentation or
normalization processes, making it difficult to identify similar windows
during the calculation of the SampEn and, consequently, the texture
patterns in the images. Even so, for segmented images, the best results
were defined by a specialist (gold standard) (Tosta, Faria, et al., 2017),
with 𝐴𝑐𝑐 rates of 98.64% (CLL × FL), 97.29% (CLL × MCL), 90.06%
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Expert Systems With Applications 202 (2022) 117238
(FL × MCL) and 88.43% (CLL × FL × MCL). In addition, considering
the results achieved from the segmentation techniques described by De
Oliveira et al. (2013), Silva et al. (2019), Tang et al. (2018), Tokime
et al. (2018), Tosta, Faria, et al. (2017), Vahadane and Sethi (2013),
and Wienert et al. (2012), the best situation to distinguish the CLL ×
FL × MCL groups was through the proposal presented by De Oliveira
et al. (2013), with 81.50% of 𝐴𝑐𝑐 rate.
Regarding the classifications obtained via normalized images, other
highlights can also be verified. The quantification of this type of image
through signature metrics indicates less expressive rates, with 𝐴𝑐𝑐
values of 82.53% (CLL × FL), 79.69% (FL × MCL), and 63.10% (CLL ×
FL × MCL), considering the model presented by Reinhard et al. (2001).
In the comparison of CLL × MCL, the best result (𝐴𝑐𝑐 rate of 78.57%)
was obtained with normalized images after applying the technique
described by Macenko et al. (2009).
In this context, the best association was defined via signature met-
rics from the original histological images with the bGWO approach.
This combination was able to distinguish the three NHL groups (CLL
× FL × MCL) with an 𝐴𝑐𝑐 rate of 98.84%. Moreover, this combination
revealed the most used metrics in this comparison. Thus, a summary of
the best association is shown in Table 6. It is noticed that the metrics 𝐴
and 𝑅 were selected in most observations 𝑚, followed by the skewness
𝑆 and maximum point of entropy 𝑃𝑚𝑎𝑥 , obtained from 𝑚 equal to 2
and 3. We believe that this is another important contribution of our
proposal, especially for expanding the quantitative capacity of SampEn,
in addition to providing and supporting new interpretations and asso-
ciations for this descriptor. In this regard, it is observed that important
approaches explore some challenges of the SampEn for quantifying and
classifying patterns, such as properly defining the parameters 𝑚 and
G.B. Rozendo et al. Expert Systems With Applications 202 (2022) 117238

Table 3
Highest 𝐴𝑐𝑐 rates achieved through SampEn signatures.
Database CLL × FL CLL × MCL FL × MCL CLL × FL × MCL
84.37% 70.21% 74.71% 59.52%
Original
bGWO bGWO GA and PSO bGWO

97.29% 95.49% 93.16% 87.86%

Segmented by specialist
PSO and bGWO SVM, K*, GA, PSO and bGWO bGWO bGWO

97.29% 93.69% 88.19% 85.54%

Segmented by De Oliveira et al. (2013)
bGWO bGWO bGWO bGWO

90.54% 93.63% 78.75% 68.60%

Segmented by Vahadane and Sethi (2013)
bGWO bGWO bGWO bGWO

93.24% 93.69% 89.44% 83.23%

Segmented by Wienert et al. (2012)
SVM, GA and PSO bGWO bGWO bGWO

95.94% 95.49% 87.57% 80.34%

Segmented by Tosta, Faria, et al. (2017)
PSO and bGWO bGWO PSO bGWO

91.89% 88.89% 83.54% 70.00%

Segmented by Tokime et al. (2018)
bGWO SVM and RF bGWO bGWO

83.78% 89.19% 78.88% 69.94%

Segmented by Tang et al. (2018)
SVM SVM bGWO bGWO

85.16% 89.19% 81.37% 73.41%

Segmented by Silva et al. (2019)
bGWO SVM bGWO bGWO

83.33% 71.91% 82.75% 64.97%

Color normalization (Reinhard et al., 2001)
bGWO bGWO bGWO bGWO

79.76% 67.65% 75.47% 58.55%

Color normalization (Macenko et al., 2009)
bGWO PSO and bGWO PSO RF

77.04% 65.92% 90.85% 53.29%

Color normalization (Khan et al., 2014)
bGWO bGWO bGWO PSO

75.21% 68.86% 74.60% 58.68%

Color normalization (Vahadane et al., 2016)
bGWO PSO RF RF

𝑟 (da Silva Senra Filho et al., 2019; dos Santos et al., 2018; Humeau-
Heurtier, 2019; Richman & Moorman, 2000; Silva et al., 2018, 2016;
Yentes et al., 2013). However, we show here that signature metrics
from SampEn are more robust for organizing multiple quantifications
and, consequently, support the pattern recognition of complex textures,
such as histological samples of NHL.
Finally, Table 7 displays a summary with the performances of the
different strategies based on SampEn to classify the three NHL groups.
Notably, signature metrics from SampEn was able to provide the most
relevant result for a classification involving CLL × FL × MCL, using
original H&E images and a reduced number of features. In addition,
this association does not require additional steps for preprocessing the
images, such as segmentation and color normalization techniques. It
is also observed that bGWO with KNN was able to provide the best
solutions (Emary et al., 2016; Mirjalili et al., 2014), including the
most relevant signature metrics to distinguish the NHL groups. The
second best association was defined from the segmented images by
a specialist and using 36 features among the 72 that constitute the
SampEn signatures. The association that indicated the lowest rate was
through the classification provided by each SampEn value, exploring
segmented images by the specialist.
5. Discussion
In this section, some additional highlights regarding the discrimi-
native ability involving the main association (signature metrics from
SampEn with original histological images and bGWO approach) are
provided. The information was organized into three sections: analy-
sis of the best association under the influence of noise; comparison
with previous works: an overview; comparison with other texture
descriptors.
5.1. Analysis of the best association under the influence of noise
The best association for distinguishing NHL groups was evaluated
by adding different types of noise, as described in Section 3.4.1. Each
test indicated the discriminative capacity of the proposed model under
adverse conditions: feature sets and images with noise levels of 20%,
40% and, 80%. Thus, the 𝐴𝑐𝑐 values are presented in Figs. 10 and 11.
Taking into account the results shown in Fig. 10, it is possible to
verify that the worst cases occurred in tests with noise levels of 80%.
Therefore, the method fails in extreme conditions: 𝐴𝑐𝑐 rates were only
50%. Also, the test considering 𝐶𝑡𝑟𝑎𝑖𝑛 × 𝐷𝑡𝑒𝑠𝑡 with noise level of 40%
was not able to provide relevant results. In this case, the 𝐴𝑐𝑐 rate was
66.15%. In these experiments, the signature metrics presented more
difficulties in distinguishing the lymphoma groups. On the other hand,
the 𝐶𝑡𝑟𝑎𝑖𝑛 × 𝐷𝑡𝑒𝑠𝑡 test with a noise level of 20% provided the highest
accuracy (92.79%). In this evaluation, the signature metrics were able
to provide an acceptable distinction. Another important fact is that the
best association was not totally affected in tests considering 𝐷𝑡𝑟𝑎𝑖𝑛 ×𝐶𝑡𝑒𝑠𝑡
and 𝐷𝑡𝑟𝑎𝑖𝑛 × 𝐷𝑡𝑒𝑠𝑡 , even with noise levels of 40%. The obtained results
were 𝐴𝑐𝑐 rates from 81.26% to 89.84%, allowing to show that the best
association is promising for distinguishing the NHL groups, even with
some adverse conditions.
Fig. 11 presents the 𝐴𝑐𝑐 rates obtained by adding RPC and WGN to
the lymphoma images classified by the best association, also in different
levels. With noise levels of 20% and 40%, the corrupted images reached
𝐴𝑐𝑐 values between 60.43% and 64.70%. With the 80% noise level,
the results are even lower, with a maximum rate of 52.40%. It is
notable that the different types of noise do not have great differences
in the obtained results, with the maximum difference of 2.94% in the
40% noise level. Therefore, the image noise types analyzed are not a

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G.B. Rozendo et al. Expert Systems With Applications 202 (2022) 117238

Table 4
Highest 𝐴𝑐𝑐 rates achieved through the classification provided by each SampEn value.
Database CLL × FL CLL × MCL FL × MCL CLL × FL × MCL
79.29% 66.80% 71.09% 59.91%
Original
K* GA, PSO and bGWO RF RF

95.94% 98.19% 77.01% 72.25%

Segmented by specialist
K*, RF, GA, PSO and bGWO RF, GA, PSO and bGWO SVM and K* K*, GA, PSO and bGWO

89.18% 91.89% 73.91% 68.20%

Segmented by De Oliveira et al. (2013)
K* RF, GA, PSO and bGWO K* K*

85.13% 89.09% 75.00% 70.93%

Segmented by Vahadane and Sethi (2013)
K* All K* K*

87.83% 90.09% 72.04% 68.78%

Segmented by Wienert et al. (2012)
K* RF, GA, PSO and bGWO K* K*

91.89% 89.18% 67.70% 63.00%

Segmented by Tosta, Faria, et al. (2017)
RF, GA, PSO and bGWO SVM, K* and RF K* K*

83.78% 92.60% 69.03% 61.18%

Segmented by Tokime et al. (2018)
SVM and K* RF, GA, PSO and bGWO K* K*

83.78% 89.18% 66.45% 58.38%

Segmented by Tang et al. (2018)
SVM and K* SVM and K* GA, PSO and bGWO K*

85.14% 89.18% 65.22% 64.74%

Segmented by Silva et al. (2019)
RF, GA, PSO and bGWO SVM and K* RF, GA, PSO and bGWO RF

78.54% 60.85% 72.03% 52.94%

Color normalization (Reinhard et al., 2001)
K* RF K* K*

75.79% 67.23% 73.18% 56.14%

Color normalization (Macenko et al., 2009)
K* K* K* K*

72.54% 62.83% 84.14% 50.00%

Color normalization (Khan et al., 2014)
K* K* All K*

76.05% 64.15% 71.82% 54.70%

Color normalization (Vahadane et al., 2016)
K* K* SVM K*

significant factor since the results are all negatively impacted. Through
this analysis, it is possible to observe that the best association is more
robust to noises added to the feature sets than to the processed images.
Fig. 11 indicates that both types of noise have more influence on
the classification than the test illustrated by Fig. 10. Considering the
three evaluations of Fig. 10, the image noise adding (Fig. 11) is only
comparable with the 80% level, with similar values in all tests, but with
lower 𝐴𝑐𝑐 values in all remaining analyses.

5.2. Comparison with previous works: An overview

Different approaches have been presented to classify NHL groups,

such as the described by Bai et al. (2019), Brancati et al. (2019),
Candelero et al. (2020), Codella et al. (2016), do Nascimento et al.
(2018), Janowczyk and Madabhushi (2016), Jiang et al. (2018), Mar-
tins et al. (2019), Meng et al. (2010), Orlov et al. (2010), Ribeiro et al.
(2018), Roberto et al. (2021, 2017), Song et al. (2012), Taino et al.
(2020), Tambe et al. (2019) and, Zhang et al. (2021). These authors
also explored the same NHL dataset, especially due to the lack of other
public sets with the technical conditions defined by National Cancer
Institute (2022) and National Institute on Aging (2022). Even with tests
based on a single dataset, each reported information has contributed
to the literature focused on related issues. On the other hand, these Fig. 10. 𝐴𝑐𝑐 rates after applying different noise levels to the feature sets.

models have not explored an approach based on SampEn signatures.

Thus, an overview of our method in comparison to the related works
is useful to illustrate the relevance of both the proposed strategy and
the results achieved. Accuracy rates reported in previously published
studies and those achieved in this paper are shown on Table 8, which
includes maximum values of 𝐴𝑐𝑐.
Regarding the studies considering NHL images (Table 8), it is pos-
sible to note that our approach may indicate results among those
available in the literature, even when we consider models based on
CNN (Bai et al., 2019; Brancati et al., 2019; Janowczyk & Madabhushi,
2016; Roberto et al., 2021; Tambe et al., 2019; Zhang et al., 2021).
Also, by taking into account that these models use different types
and totals of features to evaluate distinct properties in NHL images,
we consider the results to be rather complementary than comparable.

16
G.B. Rozendo et al. Expert Systems With Applications 202 (2022) 117238

Table 5
𝐴𝑐𝑐 rates obtained through the signature metrics from SampEn, considering the different types of images and NHL groups.
Database CLL × FL CLL × MCL FL × MCL CLL × FL × MCL
99.60% 98.72% 99.32% 98.84%
Original
bGWO bGWO bGWO bGWO

98.64% 97.29% 90.06% 88.43%

Segmented by specialist
GA GA, PSO and bGWO GA, PSO and bGWO bGWO

93.24% 90.99% 86.33% 79.76%

Segmented by Wienert et al. (2012)
PSO bGWO GA RF

91.89% 95.94% 90.06% 81.50%

Segmented by De Oliveira et al. (2013)
GA bGWO GA, PSO and bGWO GA and bGWO

91.89% 89.09% 76.25% 70.93%

Segmented by Vahadane and Sethi (2013)
GA, PSO and bGWO SVM SVM SVM

93.24% 89.18% 89.18% 74.56%

Segmented by Tosta, Faria, et al. (2017)
GA SVM bGWO SVM

93.24% 88.89% 84.18% 72.35%

Segmented by Tokime et al. (2018)
GA and bGWO SVM and RF GA and bGWO PSO and bGWO

87.84% 89.19% 87.58% 73.99%

Segmented by Tang et al. (2018)
GA, PSO and bGWO SVM RF RF

87.84% 89.18% 85.09% 77.45%

Segmented by Silva et al. (2019)
PSO and bGWO SVM RF RF

82.53% 66.80% 79.69% 63.10%

Color normalization (Reinhard et al., 2001)
GA, PSO and bGWO bGWO PSO and bGWO GA and bGWO

78.57% 78.57% 74.71% 58.28%

Color normalization (Macenko et al., 2009)
bGWO GA and PSO PSO RF

73.77% 66.81% 86.58% 54.39%

Color normalization (Khan et al., 2014)
bGWO bGWO GA and PSO bGWO

76.47% 73.11% 75.79% 54.70%

Color normalization (Vahadane et al., 2016)
bGWO GA RF PSO and bGWO

Table 6
Summary of the best association for distinguishing lymphoma classes:
CLL × FL × MCL.
𝑚 Metric Classification approach 𝐴𝑐𝑐
𝐴
𝑚=1
𝑅
𝐴
𝑚=2 𝑆
𝑃𝑚𝑎𝑥 bGWO+KNN 98.84%
𝑅
𝑚=3 𝑆
𝑃𝑚𝑎𝑥
𝐴
𝑚=4
𝑅

In addition, an important advantage of our model is that it uses a

reduced number of features to distinguish CLL × FL × MCL since the
main features have been identified. A reduced number of features can
lead to a more comprehensive solution for specialists and contribute
significantly to the community interested in the study of NHL images.
Moreover, we believe that our results obtained from the classification Fig. 11. 𝐴𝑐𝑐 rates after adding the random pixel corruption (RPC) and white Gaussian
of segmented and normalized images could encourage the development noise (WGN).
of new techniques for color normalization and segmentation of NHL
images.
2011), lacunarity (Căliman & Ivanovici, 2012; Ivanovici et al., 2009),
5.3. Comparison with other texture descriptors
and percolation (Roberto et al., 2021, 2017). These techniques use

The best association of the proposed method (signature metrics, multidimensional and multiscale approaches that are capable of pro-
original histological images, and bGWO method) was also compared viding more complete quantifications of the images, using the RGB
with other texture descriptors: fractal dimension (Ivanovici & Richard, color model as a n-dimensional space or a cubic organization. These

17
G.B. Rozendo et al. Expert Systems With Applications 202 (2022) 117238

Table 7
Summary of the different strategies used to classify the CLL × FL × MCL groups.
Composition 𝐴𝑐𝑐 Image type Number of features Classification approach
Signature metrics from SampEn 98.84% Original 10 bGWO
SampEn signatures 87.86% Segmented by specialist 36 bGWO
Classification from each SampEn value 72.25% Segmented by specialist 1 K*, PSO, bGWO and GA

Table 8
𝐴𝑐𝑐 rates of the proposed method and related works, with information about the features, types of cross-validation and classification methods.
Authors Features Number of Classifier Validation 𝐴𝑐𝑐
features
do Nascimento et al. (2018) Color channels, morphological ≈ 145 Polynomial cross-validation 10-folds 100%
features
Bai et al. (2019) CNN, texture, statistics and color ≈ 1, 188 CNN e RF hold-out 99.10%
Orlov et al. (2010) Fourier and wavelet 12–200 WND-CHARM cross-validation 8-folds 99%
Proposed method Signatures metrics from SampEn 10 bGWO cross-validation 3-folds 98.72%–99.60%
Zhang et al. (2021) Deep-learned features 2048 GA and ResNet-50 hold-out 98.63%
Jiang et al. (2018) Morphology, entropy, and other 680 Hierarchical SVM cross-validation 10-folds 97.96%
descriptors
Brancati et al. (2019) Deep-learned features ≈ 2048 CNN cross-validation 5-folds 97.67%
Tambe et al. (2019) Deep-learned features ≈ 1000 CNN hold-out 97.33%
Martins et al. (2019) Fractal dimension and lacunarity 18 Hermite polynomial cross-validation 10-folds 97.00%
with RGB and LAB color models
Song et al. (2016) Visual features 10,384 SVM cross-validation 5-folds 96.80%
Janowczyk and Madabhushi Deep-learned features 4 CNN cross-validation 5-folds 96.58%
(2016)
Roberto et al. (2017) Percolation theory 15 DECORATE cross-validation 10-folds 96.40%
Roberto et al. (2021) ResNet-50 and fractal features 2048 Double-CNN ensemble cross-validation 10-folds 95.55%
(DL+HC)
Codella et al. (2016) Color settings enhancement 200 SVM cross-validation 3-folds 95.50%
Taino et al. (2020) Fractal features, Curvelet and 73 GA with KNN cross-validation 10-folds 94.70%
Haralick
Meng et al. (2010) Visual features, wavelet, Gabor e 50 C-RSPM+WMVA cross-validation 3-folds 92.70%
LBP
Ribeiro et al. (2018) Fractal features, Curvelet 43 Decision tree cross-validation 10-folds 86.14%
sub-images and Haralick
Candelero et al. (2020) LeNet-5, Haralick, fractal features 18 K* cross-validation 3-folds 82.01%
and GA

Table 9
Classification of the CLL × FL × MCL groups using different types of texture descriptors.
Features Database Classification approach 𝐴𝑐𝑐
Signature metrics from SampEn (Proposed method) Original bGWO with KNN 98.84%
Percolation (Roberto et al., 2021, 2017) Original GA e PSO with KNN 85.54%
Lacunarity (Căliman & Ivanovici, 2012; Ivanovici et al., 2009) Segmented by specialist RF 70.52%
Fractal dimension (Ivanovici & Richard, 2011) Original K* 61.27%

descriptors were applied to quantify the histological NHL samples (orig-
inal, segmented, and normalized). The classifications were performed
with the classifiers presented in Section 3.3 and the obtained results
(𝐴𝑐𝑐 values) are shown on Table 9. It is possible to highlight that
the descriptors provided the best results from the original histologi-
cal images and also from those segmented by the specialists. These
combinations were also identified via SampEn signatures. Among the
compared descriptors, percolation (Roberto et al., 2021, 2017) was
responsible for providing the highest 𝐴𝑐𝑐 rate (85.54%) via original
images. However, this result is lower than the value achieved via
our best association (signature metrics), which further enhances the
relevance of the proposed method, even from grayscale images.
We believe that more analysis could be carried out to define
whether these results are maintained in new experiments, with different
parameters and other methods to support each indication, or even to
make adjustments and define the limits of each parameter or technique.
For instance, an integrated approach is useful to support the assessment
and ranking of concepts (Samanlioglu & Ayağ, 2021), here the main
indications (Tables 5, 7 and 9), providing a proof of concept regarding
the best combination. Also, when some methods are considered (GA,
PSO, and bGWO), it is possible to perform experiments involving
population size, parameter optimization and convergence analysis in
order to verify the impacts of these in the process of definition of the
best solution. In addition, taking into account the bGWO algorithm,
the escape probability parameter needs to be in the interval [0, 1].
However, there is a lack of published studies regarding the ideal values
for this parameter (Song, Fong, Deb, & Hanne, 2018). On the other
hand, the experiments and indications presented here were able to pro-
vide a relevant overview of our proposal compared to the consolidated
approaches commonly explored in the literature for classifying NHL
images. Moreover, considering that our strategy was able to indicate the
main associations, with a reduced number of features, the information
presented can contribute to the development of computational models
addressed to aid specialists interested in the composition of machine
learning techniques, both for NHL samples and other contexts of H&E
images.
6. Conclusion
In this paper, a new interpretation of SampEn values was accom-
plished via the exploration of parameter intervals of 𝑚 and 𝑟. These
values were considered to be SampEn signatures. To demonstrate the
relevance of using SampEn signatures, three tests were performed: a
classification based on SampEn texture signatures; classification pro-
vided by each SampEn value; and, classification using signature metrics
from SampEn, an approach that had not been explored in the literature.
This last strategy improved the quantitative capacity of SampEn, with
more interpretations and associations for this descriptor. For each

18
G.B. Rozendo et al.
test, analyses with multiple segmentation and color normalization ap-
proaches were investigated in detail. The best association was defined
by signature metrics from SampEn, original histological images and
bGWO method. This solution brings important alternatives for the
improvement of CAD systems since it provides accurate results and
does not require preprocessing steps. Only ten features were used to
successfully distinguish CLL × FL × MCL, representing a combination
with a total of descriptors considerably lower than most state-of-the-
art proposals. Therefore, signature metrics from SampEn are relevant
attributes for pattern recognition of NHL textures. The main features
were 𝐴 and 𝑅 present in most of the observations 𝑚, followed by the
𝑆 and 𝑃𝑚𝑎𝑥 metrics, obtained only from 𝑚 equal to 2 and 3.
Another important contribution is that, even in tests considering fea-
ture sets with distinct levels of noise, the best association was capable
of classifying the NHL cancer type. This fact was observed after adding
noise levels of 20% and 40%. The results obtained by our proposed
method were only hindered at noise levels of 80%. Also, it was possible
to identify that RPC and WGN have more influence on the model than
noise added to the feature sets. On the other hand, the performance
with signature metrics provided important distinctions when compared
to other techniques available in the literature. Another limitation lies in
applying the method in segmented or color normalized images, wherein
SampEn signature metrics could not perform well, especially for the
three-classes comparison, despite reaching accuracy values above 90%
on some two-classes comparison scenarios. This indicates that further
analysis of the features under pre-processed and segmented images is
required to understand this behavior. Thus, even though the proposal
failed to overcome the results of some consolidated techniques, our
study was able to indicate details of the most relevant solution with
a full understanding of the features (advantages and limits). This is an-
other important contribution to the community interested in the issues
addressed here. Therefore, we believe that the contributions provided
by this method, such as a new association of SampEn signatures and the
bGWO approach and the identification of the best features, are relevant
for the area of NHL classification. We also believe our method can be
used either as a complementary or a benchmark for future research.
In the future, we believe that investigations of different intervals
for parameters 𝑚 and 𝑟 may enhance our understanding of the limits of
the signatures in the process of classifying and recognizing the patterns
of NHL. It is important to thoroughly explore SampEn calculation
using sub-images by assessing window numbers and how different
types of distances may affect the process of verifying similarity among
windows. In addition, considering the methods explored here, such as
GA, PSO and bGWO, experiments involving population size, parameter
optimization, combinations with other classifiers (in addition to KNN)
and convergence analysis can be performed to identify the impacts
on the best associations of features and techniques for the study of
NHL groups. Finally, SampEn signatures and signature metrics can be
applied to other types of images in order to analyze their discriminatory
abilities under more conditions.
Statements of ethical approval
Ethics approval was not required for this study.
CRediT authorship contribution statement
Guilherme Botazzo Rozendo: Conceptualization, Methodology,
Software, Validation, Formal analysis, Investigation, Writing – original
draft. Marcelo Zanchetta do Nascimento: Writing – review & editing.
Guilherme Freire Roberto: Writing – review & editing. Paulo Rogério
de Faria: Formal analysis, Investigation, Data curation. Adriano Bar-
bosa Silva: Software. Thaína Aparecida Azevedo Tosta: Software,
Writing – review & editing. Leandro Alves Neves: Conceptualization,
Validation, Writing – review & editing, Supervision.
19
Expert Systems With Applications 202 (2022) 117238
Declaration of competing interest
The authors declare that they have no known competing finan-
cial interests or personal relationships that could have appeared to
influence the work reported in this paper.
Acknowledgments
This study was financed in part by the : Coordenação de Aperfeiçoa-
mento de Pessoal de Nível Superior - Brasil (CAPES) - Finance Code
001; National Council for Scientific and Technological Development
CNPq (Grants #313643/2021-0, #430965/2018-4 and #311404/2021-
9); and State of Minas Gerais Research Foundation - FAPEMIG (Grant
#APQ-00578-18).
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