international journal of refrigeration 74 (2017) 151–164

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Identification of freezer burn on frozen salmon

surface using hyperspectral imaging and
computer vision combined with machine
learning algorithm

Jun-Li Xu, Da-Wen Sun *

Food Refrigeration and Computerized Food Technology (FRCFT), School of Biosystems and Food Engineering,
Agriculture and Food Science Centre, University College Dublin, National University of Ireland, Belfield, Dublin
4, Ireland

A R T I C L E I N F O A B S T R A C T

Article history: This study explored the potential of computer vision system (CVS) and hyperspectral imaging
Received 4 August 2016 (HSI) technique covering spectral range of 900–1700 nm for identifying freezer burnt salmon
Received in revised form 13 October fillets after frozen storage. Local binary pattern (LBP) descriptor was applied for the RGB image
2016 classification. Reflectance spectra were obtained from various positions surface and pre-
Accepted 16 October 2016 treated using the standard normal variate (SNV) transformation. TreeBagger classifier was
Available online 18 October 2016 used to build classification models for recognition and authentication of the freezer burnt
flesh. The results suggested that hyperspectral discrimination performed much better than
Keywords: CVS with the correct classification rate (CCR) of 0.905 in validation and CCR of 0.945 in cross-
Atlantic salmon validation. The effective wavelengths were selected based upon the feature importance in
Hyperspectral imaging the TreeBagger model and the corresponding optimized model yielded CCR of 0.914 in vali-
Computer vision dation and 0.978 in cross-validation. Overall, the outcome suggested the capability of HSI
TreeBagger classifier for rapid categorization of damaged regions on frozen salmon.
Machine learning © 2016 Elsevier Ltd and IIR. All rights reserved.

Identification de la brûlure de congélation sur la surface du

saumon congelé en utilisant l’imagerie hyperspectrale et la
vision par ordinateur combinée avec l’algorithme
d’apprentissage automatique
Mots clés : Saumon atlantique ; Imagerie hyperspectrale ; Vision par ordinateur ; Classificateur TreeBagger ; Apprentissage automatique

* Corresponding author. Food Refrigeration and Computerized Food Technology (FRCFT), School of Biosystems and Food Engineering, Ag-
riculture and Food Science Centre, University College Dublin, National University of Ireland, Belfield, Dublin 4, Ireland. Fax: +353 1 7167493.
E-mail address: dawen.sun@ucd.ie (D.-W. Sun; Websites: www.ucd.ie/refrig, www.ucd.ie/sun.).
http://dx.doi.org/10.1016/j.ijrefrig.2016.10.014
0140-7007/© 2016 Elsevier Ltd and IIR. All rights reserved.
152 international journal of refrigeration 74 (2017) 151–164

tected food product (Schmidt and Lee, 2009). If water molecules

Nomenclature inside food keeps migrating to the surface, ice will continue to
accumulate on the surface and transfer into the air via subli-
AUC area under the ROC curve
mation. Sublimation, which is the key factor for the formation
CCR correct classification rate
of freezer burn, will continue as long as the pressure difference
CVS computer vision system
exists. As a result, some parts of meat flesh are deprived of mois-
FN false negative
ture and manifest themselves as an opaque dehydrated surface,
FOV field of view
presenting dry and shriveled appearance. An excessive desic-
FP false positive
cation is able to accelerate oxidative alterations at the surface
FPR false positive rate
of the food product. Freezer burn increases the chance for oxygen
HIS hyperspectral imaging
to come into contact with the food surface area and raises the
LBP local binary pattern
risk for oxidative reactions and structural deterioration to occur,
OOB out-of-bag
which can irreversibly change food color, texture and flavor (Evans,
PCA principal components analysis
2008). Freezer burn usually appears as grayish-brown leathery
PCs principal components
spots, papery or cottony edges and a dry texture on frozen food.
RGB red green blue
Because of unpleasant flavor, it is highly recommended to iden-
ROC receiver operator characteristics
tify and remove the parts affected by freezer burn.
ROIs regions of interest
Traditional estimation of freezer burn relies on manual as-
SD standard deviation
sessment by trained evaluators using standardized methodology,
SNR signal to-noise ratio
which is time-consuming, tedious and subjective. Computer vision
SNV standard normal variate
system (CVS) (Du and Sun, 2005; Jackman et al. 2009; Wu and
TN true negative
Sun, 2013c) is a modern and efficient approach for monitoring
TP true positive
quality traits in salmonids including color (Misimi et al., 2007;
TPR true positive rate
Quevedo et al., 2010), shape and size (Misimi et al., 2008a), firm-
ness (Quevedo and Aguilera, 2010), and fat content (Stien et al.,
2007). However, CVS normally has no ability to assess the in-
ternal quality attributes because it is conducted in visible
1. Introduction wavelength range (Wu et al., 2012). Therefore, hyperspectral
imaging (HSI) has recently emerged as a versatile and promis-
Farmed salmon, as a gourmet fish with high nutritional value, ing analytical tool in food quality control (Sun, 2010; Barbin et al.,
is now marketed either as fresh whole fish, fillets or packaged 2012; Elmasry et al., 2012; Feng and Sun, 2012; Barbin et al., 2013;
as frozen or smoked products. Clearly, as other techniques such ElMasry et al., 2013; Feng and Sun, 2013; Feng et al., 2013;
as cooling (Sun, 1997; Sun and Brosnan, 1999; Hu and Sun, 2000; Kamruzzaman et al., 2013; Wu and Sun, 2013d; Liu et al., 2014).
Sun and Wang, 2000; Mc Donald and Sun, 2001; Wang and Sun, By integrating conventional imaging and spectroscopy into one
2002a, 2002b; Sun and Hu, 2003; Wang and Sun, 2004; Zheng and system, HSI is capable of collecting both spatial and spectral in-
Sun, 2004) and drying (Kiani et al. 2011), freezing (Sun and Woods, formation from a sample (Kamruzzaman et al., 2015). In terms
1994; Cui et al. 2008) is an effective food preservation tech- of the applications in salmon, HSI technique has recently been
nique, and thus is the safest and most commonly-used means successfully used for determination of fat, moisture (Zhu et al.,
to maintain salmon quality for long-term storage (Kilincceker 2014), and sodium (Huang et al., 2003), measurement of caloric
et al., 2009; Rodriguez-Turienzo et al., 2011).The market for frozen density (Xu et al., 2016a), and evaluation of microbial spoilage
food has remarkably expanded worldwide over the last 50 years in salmon flesh (Wu and Sun, 2013b). Nevertheless, to the best
(Tee et al., 2014). However, fish qualities including color, texture, of our knowledge, no research has been reported to detect freezer
and flavor may decline during long periods of frozen storage burn on the frozen salmon using neither computer vision system
(Arannilewa et al., 2005). Most importantly, freezer burn will occur nor hyperspectral imaging technique. Hence, the objective of this
and cause extreme toughening, discoloration of the surface and study was to investigate the suitability of using CVS and HSI
the development of rancidity in the affected area if fish is not technique in the long-wave near infrared range (900–1700 nm)
appropriately wrapped during frozen storage. This surface des- for detection of freezer burn on frozen salmon slices.
iccation defect results from sublimation which is very possible
to occur in situations where frozen food are preserved without
an adequate moisture barrier packaging (Kaess, 1961). 2. Materials and methods
Sublimation from the surface region of the food happens when
water pressure over the surface of the food product is higher 2.1. Salmon sample preparation
than the vapor pressure of water in the air (Ashby et al., 1973).
In order to equilibrate with the vapor pressure of the environ- A total of 60 conventional farm-raised Atlantic salmon (Salmo
ment, the ice will not melt but escape into the air directly from salar) fillets were purchased from Marine Harvest Ireland
solid phase to vapor state. Meanwhile, water vapor in the air Company. Salmon samples were reared in the sea with the tem-
has an overall tendency to accumulate directly as ice on the re- perature of 15.4 °C before they reached commercial size, between
frigeration coil since the temperature of the coil is always lower 3 kg and 5 kg, which normally takes between 18 and 24 months.
than the air in the refrigerator system, which provides the driving After harvesting, fresh salmon were beheaded, filleted and then
force for constant loss of water from the surface of the unpro- transported to laboratories of Food Refrigeration and Comput-
 international journal of refrigeration 74 (2017) 151–164
erized Food Technology (FRCFT), University College Dublin (UCD),
Ireland. All the fillets were placed on ice and packaged with a
big plastic barrier during transport and the temperature inside
the storage area was maintained at −1 °C to make sure the quality
would not deteriorate. Each fillet was about 100 grams in dif-
ferent sizes and it was guaranteed that every fillet was cut from
different fish.The salmon fillets were subsampled into a cuboid
shape with approximate sizes of 6.0 cm × 6.0 cm × 3.0 cm
(length × width × thickness) to acquire 120 subsamples, which
were obtained from different locations of salmon fillets. Samples
were randomly divided into three groups with equal numbers:
the first group samples were packed in sealed freezer bags, the
second group was wrapped in plastic bags and the third group
samples were unwrapped on a tray. All three groups were stored
in a chest freezer with the temperature of −18 °C for two weeks
and then thawed overnight at controlled refrigerated condi-
tions (1 ± 1 °C). Each sample was first scanned by CVS and then
by the HSI system and it was guaranteed that no obvious liquid
drops were found on the salmon surface during scanning.
It was believed that first group samples packed in sealed freezer
bags were of good quality, while samples from the second group
were likely to develop some freezer burnt regions and the third
group samples had a strong possibility to have many freezer burnt
areas. In this study, freezer burn regions were determined by
using the traditional manual assessment. The visual observa-
tions included the evident color change, the appearance of the
whole blemished areas or leathery spots, the papery or cottony
edges and a dry texture on the meat surface. In addition, pre-
vious research had demonstrated that weight loss was an
important parameter to evaluate the occurrence of freezer burn
on meat (Kaess and Weidemann, 1967). Therefore, percentage
of weight loss of individual fillet was also measured using a mass
balance to help define a freezer burnt sample.
2.2. Computer vision system
The computer vision system (CVS) used in our study, described
by Valous et al. (2009), consists of three main components: a light-
ing system consists of a cold light unit (Kaiser Fototechnik,
Germany) with four light banks, each one fitted with one 36 W
daylight fluorescent lamp, a color digital camera (Canon
PowerShot A75, Canon, USA), and a computer (Dell Computer
Corporation, USA) installed with a remote capture software
(version 2.7.5.27, Canon, USA) to acquire and visualize the digi-
tized images. The entire system was housed in a black wooden
box with internal walls painted black to alleviate the external
light and reflections (Saenz et al., 2005). Four fluorescent lamps
were arranged in the form of a square, at an angle of approxi-
mately 45° in relation to the sample. Salmon samples were
illuminated in a light blue background similar to the color of
conveyor belts sometimes used in fish processing plants (Misimi,
2007). An object of known size demonstrated that an image area
of 18 × 18 pixels corresponded to 1 mm2 in reality. Calibration,
based on the method proposed by Leon et al. (2006), was per-
formed to ensure that the salmon images taken from the digital
camera were true representations of the fish in the scene when
it comes to their real-world dimensions and color (Misimi et al.,
2008b). All the algorithms for image pre-processing, segmen-
tation, feature extraction, feature selection and classification of
153
salmon were performed in Matlab 8.5 R2015b software (The
Mathworks Inc., MA, USA).
2.3. Hyperspectral imaging system
Spectral images of the prepared samples were acquired in the
reflectance mode by employing a laboratory-based line-
scanning hyperspectral imaging system covering the wavelength
region of 900–1700 nm. The specific description of this system
can be found elsewhere (Xu et al., 2016b). The spectrograph with
a spectral increment of approximately 3.34 nm between con-
tiguous bands had a fixed-size internal slit to denote a field
of view (FOV) for the spatial line (horizontal pixel direction).
Each salmon sample was individually placed on the transla-
tion stage and then conveyed to the FOV of camera with a speed
of 2.7 cm s−1. The acquired three-dimensional hyperspectral
images were stored in raw format and then exported to Matlab
8.5 R2015b software (The Mathworks Inc., MA, USA) for sub-
sequent processing. Since HSI system collects the detector signal
intensity, rather than actual reflectance spectra, the original
images were calibrated into the reflectance images based upon
the standard calibration procedure suggested by Wu et al. (2012).
2.4. Identification of freezer burn by CVS
2.4.1. RGB image preprocessing
A binary mask can yield an image containing only the salmon
sample and avoid any interference from the background. The
background was removed from the RGB image by subtracting
a G (Green) channel from R (Red) channel followed by a simple
thresholding. Afterwards, morphological operations were also
used to remove some defects that were falsely classified as the
background in the binary mask (Mery et al., 2013).
After removing the background, the masked RGB image was
then converted to the grayscale image. Wiener filtering tech-
nique was implemented to the grayscale image to reduce the
influence of the noise in the feature extraction and selection
step. In detail, the wiener2 function was applied in Matlab en-
vironment using neighborhoods of size 5-by-5. This function
employs a pixel wise adaptive Wiener method based on sta-
tistics estimated from a local neighborhood of each pixel as
shown in Eq. (1), Eq. (2) and Eq. (3) as follows:
1
μ= ∑ a (n1, n2 )
NM n1,n2 ∈η
(1)
and
1
σ2 = ∑ a2 (n1, n2 ) − μ 2
NM n1,n2 ∈η
(2)
where η is the N-by-M local neighborhood of each pixel in the
image, while μ and σ2 are local mean and variance around each
pixel (n1,n2), respectively. Adaptive Wiener method then pro-
duces a pixel wise Wiener filter using these estimates,
σ 2 − v2
b (n1, n2 ) = μ + (a (n1, n2 ) − μ ) (3)
σ2
where v2 is supposed to be the noise variance. If not given, wiener2
calculates the average of all the local estimated variances.
154 international journal of refrigeration 74 (2017) 151–164
2.4.2. Feature extraction
Local binary pattern (LBP) is a simple but efficient texture op-
erator. It aims to label the pixels of an image by thresholding
the neighbor pixels with the center pixel value and records the
result as a binary number. Because of its discriminative power
and computational simplicity, LBP texture operator has been
widely used in various applications, including texture classi-
fication and segmentation, image retrieval and surface
inspection (Chang-yeon, 2008). The original LBP operator (Ojala
and Pietikäinen, 1997) was built by first dividing the exam-
ined window into 3 × 3 neighborhood followed by thresholding
the value of the neighbor pixels compared with center pixel
and then summing up the obtained values of pixels which had
been multiplied by the given binomial weights. Given a central
pixel in the image, a pattern number is calculated by compar-
ing its value to its neighbors:
p−1
LBP ( xC, yC ) = ∑ s (gp − gC ) 2 p (4)
p= 0
where gC means the grey value of the center pixel (xC, yC), gp
corresponds to the grey values of the P neighborhood pixels
involved, while the function s(x) is determined as follows:
⎧1 x ≥ 0
s (x ) = ⎨ (5)
⎩0 x < 0
One extension to the original operator was the application
of neighborhoods with different sizes (Ojala et al., 2002). Any
radius and number of pixels in the neighborhood was allowed
by using a circular neighborhood and bilinearly interpolating
values at non-integer pixel coordinates. Another important ex-
tension of the LBP operator is the so-called uniform pattern
(Heikkila and Pietikainen, 2006) to reduce the length of the feature
vector, which was accomplished by implementing a simple
rotation-invariant descriptor.This extension was available because
of the fact that some binary patterns have higher occurrence
rates in texture images than others. The LBP pattern is consid-
ered as uniform if it has limited transition or discontinuities (at
most two) in the circular binary presentation (Ojala et al., 2002).
For instance, the patterns 00000000 (0 transitions), and 11000111
(2 transitions) are qualified while the patterns 11001101 (4 tran-
sitions) and 00101011 (6 transitions) are not. It was believed that
only “uniform” patterns are fundamental patterns of local image
texture. In practice, the mapping from LBPP,R to LBPP,Ru2 (the sub-
script represents having P neighbors on a circle of radius R and
the superscript u2 stands for using only uniform patterns and
labeling all remaining patterns with a single label), which has
P ∗ ( P − 1) + 3 distinct output values, is used with a lookup table
of 2P elements (Guo et al., 2010).
In this study, 200 regions of interest (ROIs) were extracted
from RGB images to represent the normal flesh and 200 ROIs
were selected to stand for freezer burnt flesh from 120
subsamples. Each ROI was of the same size with 100 × 100
pixels. The uniform LBP16,1 operator was implemented to extract
texture feature descriptor of image as shown in Fig. 1. Firstly,
RGB image of the size 100 × 100 was converted into grayscale
image and then divided into neighborhoods in which each pixel
was compared with the center pixel value to obtain a binary
number. Histogram of the frequency of each number occur-
ring (i.e., each combination of which pixels are smaller and
which are greater than the center) was computed and formed
a multi-dimensional feature vector, in our case, 243-dimensional
feature vector was successfully obtained. LBP histogram was
subsequently normalized and used as representation of one
image sample. 180 image samples with the size of 100 × 100
was utilized as the training dataset, therefore, the LBP descrip-
tor would be the size of 180 × 243 and it now can be processed
to train machine learning algorithm.
2.5. Identification of freezer burn by HSI
2.5.1. ROIs identification
Background removal is a fundamental step to separate the
salmon sample from the background in hyperspectral image,
as subsequent extracted data are highly based on the preci-
sion of this process (Kamruzzaman et al., 2012). In this work,
background was eliminated from the hyperspectral image by
subtracting a low-reflectance band from a high-reflectance band
followed by a simple thresholding. Fig. 2 displays the spectral
difference between background and salmon sample. As can be
seen, two wavelengths should be selected, one with high re-
flectance values (Band 1 of 1080 nm) and the other with low
reflectance values (Band 2 of 1420 nm), and subtracted from
each other to obtain a grayscale image. The obtained grayscale
image had uniform gray level and no extreme pixel can be ob-
served. All the pixels in background were set to approximately
0 because each spectrum of background remains constant along
the whole wavelength region. As a result, thresholding value
of 0.01 was obtained and a binary mask was generated. After-
wards, 200 ROIs with the specified size of 10 × 10 were extracted
from the masked image to represent normal flesh and another
200 ROIs to denote freezer burnt flesh.
2.5.2. Spectra pre-processing
180 wavelength variables in the range of 900–1700 nm were re-
tained after eliminating signal noise at the beginning and end
of the spectral region (Moscetti et al., 2015). Standard normal
variate (SNV) pre-processing was applied to remove both ad-
ditive and multiplicative effects in the spectra (Pu et al., 2015).
In SNV, each spectrum is first centered and then scaled by the
corresponding standard deviation (Barnes et al., 1989). In this
work, the spectrum of each pixel was processed by using SNV
and the spectra of each 100 pixels were then averaged to rep-
resent each ROI. The extracted spectral data were arranged in
a spectral matrix (X) where the columns of this matrix repre-
sent the wavelengths and the rows stand for the samples.
Among 400 ROIs, 180 were randomly selected as the calibra-
tion dataset and the remaining 220 as the validation set.
2.5.3. Principal components analysis
It is imperative to explore and understand the structure of the
hyperspectral image in order to select the proper methods for
dealing with the final aim. Among all the tools for multivari-
ate exploration, principal components analysis (PCA) is the most
frequently-used one for data compression and exploratory
analysis (Davies, 2005; Gemperline, 2006). In hyperspectral
imaging, PCA enables dimension reduction by using an or-
 international journal of refrigeration 74 (2017) 151–164 155

Fig. 1 – Key steps for the whole procedure of model development by CVS.

thogonal transformation to convert many spectral variables

which are possibly correlated variables into a smaller number
of linearly uncorrelated variables called principal compo-
nents which capture the maximum variation in the data (Gowen
et al., 2009). PCA makes the transformed vectors uncorrelated
and orthogonal thereby removing co-linearity problem
(Alexandrakis et al., 2012). PCA was applied to a calibration
dataset comprising of 90 normal spectra and 90 freezer burnt
spectra to explore the spectral difference between these two
classes and the possibility of classifying them.

2.6. TreeBagger classifier

Machine learning is actually a subfield of computer science and

it generally refers to the process of first building a model from
the training set and then making data-driven predictions on
unknown data based on the developed model. Machine learn-
ing is applied in a variety of computing tasks especially when
designing and programming explicit algorithms are unsuit-
Fig. 2 – Spectral difference between background and able. Supervised learning is a sub-field of machine learning in
samples. which a set of labeled training data are used to train the
156 international journal of refrigeration 74 (2017) 151–164
algorithm by developing a statistical model which can be
applied to unknown data to predict the classification label (Yu
et al., 2012). In this work, the extracted feature data were ana-
lyzed and modeled using TreeBagger supervised machine
learning function in Matlab 8.5 R2015b software (The Mathworks
Inc., MA, USA). In our case, the TreeBagger classifier employs
random forests with bootstrap aggregation to train the algo-
rithm. Random forests algorithm is an ensemble learning
method that is developed first by using a collection of deci-
sion trees and then combining the predictions to yield more
accurate classification result (Cutler et al., 2007). Random forests
attempt to reduce the variance by averaging multiple deep de-
cision trees and trained on different parts of the same training
set. This also helps avoid overfitting which is likely to appear
if only one decision tree is used. The algorithm begins with the
selection of many bootstrap samples from the dataset and those
not occurred in a bootstrap sample are called out-of-bag (OOB)
observations. Every decision tree is grown on an indepen-
dently drawn bootstrap replica of input data and the prediction
is computed by using the ensemble average. Meanwhile, boot-
strap aggregation (or bagging) is also designed to improve the
stability and accuracy of machine learning algorithms because
it reduces the variance of the model, without raising the bias.
Bootstrap sampling is a method of de-correlating the trees by
introducing them different training sets since trees grown in
the same data set would be sensitive to the noise within that
data and be correlated (Szymanska et al., 2016). Therefore, the
variance and the correlation between trees are both de-
creased by bagging and averaging the results obtained for each
tree. An optimal number of trees is normally obtained from
inspecting how the out-of-bag error changes with accumula-
tion of trees. In general, using more decision trees always
indicates a more reliable estimate obtained from out-of-bag
predictions. However, the improvement decreases as the
number of decision trees increases, which means at a certain
point the benefit in prediction performance from using more
trees will be lower than the cost in computation time for learn-
ing these additional growing trees. In this implementation,
N = 50 decision trees were first empirically determined on a
training set with the feature set as a predictors and the actual
label as the response. Then, the change pattern of the en-
semble errors against the accumulation of trees would be
inspected to further evaluate whether 50 growing trees were
sufficient or not. The TreeBagger algorithm can also rank the
importance of variables used to build classification model.
Thereby, some uninformative features or wavelength vari-
ables can be eliminated from the model to reduce the
computation time and improve accuracy.
2.7. Model performance evaluation
For the TreeBagger classification model, accuracy could be quan-
tified using the out-of-bag classification errors versus number
of grown trees. In our situation, model was mainly statisti-
cally assessed by using correct classification rate (CCR),
sensitivity and specificity for a 10-fold cross-validation and vali-
dation set (Amigo et al., 2015):
TP
Sensitivity = (6)
TP + FN
TN
Specificity = (7)
FP + TN
In the equations, TP and TN refer to true positive and true
negative, representing positive and negative pixels that are cor-
rectly identified, respectively. On the other hand, FP and FN
denote false positive and false negative, respectively. In sta-
tistics, the receiver operator characteristics (ROC) curve, which
is a graphical plot, is also widely used to evaluate the perfor-
mance of the classification model as its discrimination threshold
is different (Lorente et al., 2013). Technically, the ROC curve is
generated by plotting the true positive rate (TPR) against the
false positive rate (FPR) at different possible threshold set-
tings of a diagnostic test. A closest point in the ROC curve to
the top left corner of the chart or coordinate (0, 1) usually results
from an optimal threshold. A random guessing would lead to
a point along a diagonal line from the left bottom to the top
right corner. Area under the ROC curve (AUC) can also be applied
to compare general classification performances because of its
statistical property: the AUC of a classification model is equiva-
lent to the probability that the classification model would rank
a randomly selected positive instance higher than a ran-
domly chosen negative one (Ariana et al., 2006).
3. Results and discussion
3.1. RGB images of normal and freezer burnt flesh
Representative color images of the salmon ROIs under inves-
tigation in this study are presented in Fig. 3. As shown, the
appearance of discoloration is observed in the freezer burnt
flesh, together with the dehydrated surface and leathery texture.
Salmon is a fat-rich fish with a large proportion of lipids con-
gregated in white stripes of connective tissue (myocommata),
segmenting the red-colored muscle tissue in vertical blocks and
presenting a zebra-like appearance (Borderías et al., 1999). The
heterogeneous color appearance makes the classification of
freezer burnt flesh based on color difference practically im-
possible. Weight loss (percentage of initial fillet weight)
for the second group samples (mean ± standard deviation
(SD) = 4.78 ± 4.13%) were observed to be slightly higher than that
of the first group (mean ± SD = 3.59 ± 2.86%). Nevertheless, for
salmon fillets in the third group, which were unprotected during
the whole frozen storage, a much higher weight loss of
11.60 ± 1.28% was observed. There was significant difference
in weight loss between the first and third groups (p < 0.01).
3.2. Classification model of CVS
From the collected training sets, LBP histograms were ob-
tained and used as inputs for the TreeBagger classifier. Table 1
shows the performances of the classification models applied
in the training and validation sets in terms of sensitivity, speci-
ficity and correct classification rate (CCR). Sensitivity is the
proportion of samples detected as positive that actually are posi-
tive, whereas the specificity is the proportion of negative sample
objects that are correctly identified. As can be seen, the
 international journal of refrigeration 74 (2017) 151–164 157

Fig. 3 – RGB images of 10 normal flesh and 10 freezer burnt flesh.

TreeBagger model (Model I) yielded a reasonable result with
CCR of 0.839 for cross validation and 0.841 for an external vali-
dation set. The TreeBagger algorithm can also quantify the
relative OOB importance of each feature as displayed in Fig. 4a.
In general, irrelative or informative features should be elimi-
nated to improve processing speed. Therefore, features whose
OOB feature importance were not higher than 0.38 were
removed and only five feature variables were finally retained.
Having selected the most important features, a simplified clas-
sification model was subsequently developed. As shown in
Table 1, it is observed that a better performance in cross vali-
dation was achieved by simplifying the classification model
(Model II) with a higher CCR of 0.850. The confusion matrix in
Table 2 presents that 19 freezer burnt samples are misclassified
into normal group and 23 normal samples are wrongly iden-
tified as damaged class.
The detailed comparison between full feature model (Model
I) and reduced model (Model II) are presented in Fig. 4. As shown
in Fig. 4b, the OOB classification error significantly decreases
from approximately 0.17 built from the full features to less than
0.15 of using 5 features. Additionally, it indicates that 50 clas-
sification trees were more than enough to obtain a stable OOB
classification error no matter for the full feature model or the
reduced one.

Table 1 – Model performance for identification of freezer burnt salmon flesh.

Model Variable Cross validation Validation
number
CCR Sensitivity Specificity CCR Sensitivity Specificity
I 243 0.839 0.822 0.856 0.841 0.836 0.846
II 5 0.850 0.867 0.833 0.809 0.791 0.827
III 180 0.945 0.946 0.936 0.905 0.873 0.936
IV 5 0.978 0.989 0.967 0.914 0.873 0.955
CCR: Correct classification rate.
158 international journal of refrigeration 74 (2017) 151–164

vibrations of the molecular chemical bonds, such as O—H, C—H

Table 2 – Confusion matrix obtained from simplified
and N—H (Ghosh and Jayas, 2009). The presence of water in
model using CVS.
the salmon flesh showed two absorption bands at 980 nm and
Predicted Predicted as
1450 nm due to O—H stretching second and first overtones (Xu
as normal freezer burnt
et al., 2016b) in Fig. 5. Besides, the other absorption peak around
Normal flesh 87 23 1200 nm was ascribed to the C—H stretch second overtone of
Freezer burnt flesh 19 91
fat (Wu and Sun, 2013a). It is also observed that the spectral
reflectance curves of samples are quite smooth with similar
trends across the whole tested wavelength region. However,
3.3. Spectral features of salmon chops a remarkable difference in the magnitudes of spectral reflec-
tance value between the normal and damaged flesh can be
The mean and standard deviation spectra of the normal and noticed. Most of freezer burnt samples exhibited higher re-
damaged samples from the calibration set are presented in Fig. 5 flectance values in the whole wavelength range of 964–
with the title of Raw Spectra. It was widely believed that the 1564 nm. This was caused by the changing of the major
spectral data extracted from the pixels within the ROI con- chemical compositions in salmon fillets during the frozen
tains the information of chemical molecules which was process. Meanwhile, SNV pre-treated spectra of calibration set
associated with the chop’s composition (Wu and Sun, 2013b). are also demonstrated in Fig. 5. It is noticed that the abscissa
Some broadband peaks occurring in the near infrared region (y-axis) has changed after SNV pre-processing. The spectra now
can be well explained by the overtone and combination are centered on the zero axis, resulting from the mean sub-

Fig. 4 – TreeBagger classifier performance based on CVS data.

 international journal of refrigeration 74 (2017) 151–164
Fig. 5 – Top: Spectral profiles of normal and freezer burnt salmon samples. Bottom: PCA model performance in terms of
score plot and loadings.
traction. It is also obvious that SNV gives tighter groupings of
the measurements.
3.4. PCA exploration result
After being pre-processed with SNV and mean center, the cali-
bration set of hyperspectral data was used to build PCA
unsupervised model to compress spectral information into a
small number of PC scores. The total amount of variance ex-
plained by the first two PC scores was higher than 97%, denoting
that the main differences between normal and defected flesh
are very possibly within these two PCs. The score plot of PC 1
against PC 2 for the calibration set is shown in the bottom row
of Fig. 5. It can be seen that all sample points are generally clus-
tered into two groups based on their first two orthogonal PCs.
Consequently, it can be concluded that normal and damaged
classes are separable due to the reasonable variation in the spec-
tral patterns of the examined samples. Additionally, normal
samples are found to be more dispersed and located at a dif-
ferent region in this particular two-dimensional space, while
freezer burnt samples seems to obey a linear trend in the PC
1-PC 2 score plot. Also, the loadings for the PC 1 and PC 2 (Fig. 5)
suggest that the main source of difference is centered around
1220 nm, which presents the highest negative loading values
on PC 1 and the highest positive values on PC 2. Although PCA
is not a supervised classification model, with PCA we can
explore the structure of hypercube, study data variance and
acquire some clues about the classifier which will be used
159
afterwards (Cen et al., 2014). Overall, the results of PCA indi-
cated that the tested samples could be separated into two
distinct categories based upon their spectra, which facili-
tates building a robust calibration model to identify freezer
burnt area in salmon flesh.
3.5. Classification model of HSI
In order to estimate a boundary to separate the clusters of
freezer-burnt and normal spectra, the TreeBagger classifier was
applied. To create a calibration model, X matrix with the ex-
tracted reflectance spectral data within full spectral range (180
independent wavelength variables) and the corresponding Y
matrix containing the belonging identity for each class, are
needed. The statistical results in the calibration and valida-
tion sets are also revealed in Table 1. As shown, the TreeBagger
classifier built with the full spectral range exhibited good per-
formance to identify damaged salmon samples with the CCR
of 0.945 in cross-validation and 0.905 in prediction. More in
detail, the sensitivity for this model developed based on the
spectral data is 0.946 in cross-validation and 0.873 in valida-
tion compared with 0.822 in cross-validation and 0.836 in
validation for the CVS method. Additionally, the specificity ob-
tained from this model (Model III) in cross-validation and
validation is observed to be much higher than that in the CVS
model (Model I). Consequently, compared with CVS, model built
from hyperspectral data can detect a greater number of false
alarms, but at the same time detect a greater number of real
160 international journal of refrigeration 74 (2017) 151–164
Fig. 6 – TreeBagger classifier performance based on HSI data.
flaws, leading to a higher accuracy in Model III. The detailed
model performance exhibition is presented in Fig. 6 with OOB
classification error and ROC curves included.
Since hyperspectral images are acquired with redundant in-
formation among contiguous wavelengths, HSI usually finds
it very difficult in high speed data acquisition and processing
(ElMasry et al., 2012). Thereby, selecting some effective wave-
lengths carrying the most valuable information to build a
simplified spectral model will generate equally or more sat-
isfactory prediction results (Cheng and Sun, 2015). In this study,
five wavelength variables were selected using the arbitrary
metric of out-of-bag feature importance. Fig. 6a shows five in-
dividual wavelengths obtained (1014, 1188, 1225, 1228 and
1406 nm), which were recognized as the effective wave-
lengths for further classification. It is noticed that the
wavelength of 1406 nm contributes most to the model devel-
opment in terms of feature importance due to the fact that
one of the major water absorption features is centered near
1450 nm because of first overtone of the O—H stretching band
(Jacquemoud and Ustin, 2003). In addition, Büning-Pfaue (2003)
conducted research to investigate the changes in spectra of
sliced pear flesh during a process of dehydration. It was found
that the sharp absorption band at the wavelength of 1406 nm
decreased in intensity, along with the process of dehydration
(Büning-Pfaue, 2003). The selection of 1225 and 1228 nm is in
good agreement with the loading plots in PCA model where
it indicates that the main source of spectral difference is cen-
tered on 1220 nm. In addition, Clevers et al. (2008) addressed
that 1220 nm was the combination absorption wavelength in
pure water. In most cases, many spectral variables mainly
include irrelevant or redundant information, or have low signal-
to-noise ratio (SNR), thus, the inclusion of these wavelengths
will not improve the model performance. The removal of these
irrelevant variables is able to promote the results in terms of
accuracy and robustness (Wu et al., 2009).
As shown in Table 1, the classification ability of the opti-
mized model (Model IV) possessed better performance than
the model developed by using the whole spectral range. There
is a remarkable improvement observed in CCR from 0.945 to
0.978 for cross-validation and from 0.905 to 0.914 for valida-
tion. As displayed in Fig. 6b, the OOB classification errors for
the reduced model were less than 0.05 after 10 grown trees.
It also demonstrates that 50 decision trees were quite suffi-
cient to reach a constant OOB error for both the full wavelength
model and the simplified one. Good classification ability of this
model (Model IV) is also confirmed by ROC curves in Fig. 6.
Moreover, according to the confusion matrix in Table 3, of the
220 prediction samples in validation set, 201 are correctly iden-
tified. It is important to point out that only 5 freezer burnt
samples are wrongly classified as normal samples.
 international journal of refrigeration 74 (2017) 151–164 161

3.6. Classification maps

Table 3 – Confusion matrix obtained from simplified
model using HSI.
The reduced model (Model II) built by using CVS was applied
Predicted Predicted as
to generate classification maps as shown in Fig. 7a. Different
as normal freezer burnt
colors in classification map denote that the detected freezer
Normal flesh 96 14 burnt regions are disconnected. As indicated, the tested samples
Freezer burnt flesh 5 105
are cut from different parts of a whole salmon and the freezer
burnt regions distribute randomly, some are obvious while the

Fig. 7 – Classification map for TreeBagger classification method applied to (a) CVS data and (b) HSI data. Different colors in
classification map refer to disconnected freezer burnt regions.
162 international journal of refrigeration 74 (2017) 151–164
others are difficult to discern with naked eyes based upon only
the images (the first sample in Fig. 7a). It is important to note
that the boundary of the sample was added to each classifi-
cation map to highlight the location of the real freezer burnt
candidate region. The results show that the model performed
reasonable well for identification of freezer burnt salmon fillets
with some acceptable false alarms detected.
Concerned with hyperspectral imaging, the simplified model
with only 2.8% data of the original matrix was executed to vi-
sualize prediction map as displayed in Fig. 7b. Firstly, each
hypercube was unfolded into a two-dimensional matrix where
each row represented the spectrum from each pixel and each
column referred to a certain wavelength variable. The TreeBagger
model (Model IV) was then used to classify each pixel into
normal or freezer burnt categories. The obtained matrix con-
tains only “0” if the pixels was classified as normal sample or
“1” if it was recognized as freezer burnt flesh. This matrix was
finally transformed to generate a classification map. Mean-
while, false color images obtained from PCA score images are
also shown for comparison. The first three score images in PCA
model (each of the score images is a grey-level image) which
contained most of information were assigned to red, green and
blue channels to form a false RGB image. In the false RGB image,
any reddish, greenish or bluish colors observed stands for the
first, second and third principal components, respectively. It
can be seen that most of the freezer burnt regions in the image
appear reddish, implying high values of the red channel (PC
1), which agrees with the score plot in the Fig. 5 where the most
of the freezer burnt samples intend to have positive scores on
PC 1.
Tested samples 4 and 5 were obtained from the third group
without any protection during frozen storage, while samples
1, 2 and 3 were from the second group wrapped with plastic
bags. Visually, samples 4 and 5 have suffered from severe freeze
damage with most of surface covered with freezer burn regions.
Both CVS and HSI were able to detect big freezer burn areas
in these two samples, yet CVS failed to accurately predict the
edge regions. With respect to sample 3, better result was ob-
served by using HSI because CVS misclassified some central
pixels into the damaged class and at the same time the true
freezer burnt pixels at the bottom edge were not fully identi-
fied. As for samples 1 and 2, it was found that HSI technique
could be affected by white strips in salmon fillets. In spite of
that, HSI still outperformed CVS in terms of the accuracy to
identify freezer burn regions in sample 1. The true freeze
damaged regions were distributed along the top and right edges
of sample 1. Obviously, too many pixels were wrongly classi-
fied into the damaged class by using CVS. Overall, the
performance of hyperspectral imaging in terms of prediction
maps was promising: the majority of pixels from the freezer
burnt region were correctly identified, even at early stages when
the effect of freezer burn was not clearly visible (the first sample
in Fig. 7a).
4. Conclusions
In this work, the feasibility of using CVS and HSI technique
in the long-wave near infrared range (900–1700 nm) was in-
vestigated for rapid and non-destructive detection of freezer
burn in frozen salmon slices. It was found that freezer burnt
salmon flesh possess the characteristic of the appearance of
discoloration, dehydrated surface and leathery texture. Clas-
sification results demonstrated the greater potential of
hyperspectral imaging compared with conventional RGB
imaging, better performances were observed in both cross-
validation and validation. TreeBagger algorithm was testified
as a robust bagging method which can produce high classifi-
cation accuracy. Five effective wavelengths (1014, 1188, 1225,
1228 and 1406 nm) were selected based on the feature impor-
tance and the best performance was achieved by using this
reduced model with the CCR of 0.978 for cross-validation and
0.914 for validation. In conclusion, hyperspectral imaging com-
bined with machine learning algorithm have great potential
for identification of freezer burn in frozen salmon fillets before
visible damage was evident.
Acknowledgments
The authors would like to acknowledge the UCD-CSC Schol-
arship Scheme supported by University College Dublin (UCD)
and China Scholarship Council (CSC) for financial support of
this study.
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