7574 IEEE TRANSACTIONS ON INSTRUMENTATION AND MEASUREMENT, VOL. 69, NO.

10, OCTOBER 2020

Turbidimetry by Image Degradation Analysis

Alejandro J. Gimenez , Francisco Javier Medellín-Rodríguez , Lydia M. Contreras-Martinez ,
José Mauricio López-Romero , Isaac C. Sanchez , Francisco Villaseñor-Ortega ,
and Gabriel Luna-Bárcenas

Abstract— Here, we describe a method to measure turbidity
in liquids by probing a graphic pattern by optical transmission.
Data are collected by a video camera, and a mathematical
algorithm is used to correlate image loss with turbidity. We report
high-resolution turbidity measurements of nanoparticle colloids
to assess the proposed methodology. Measurements are differen-
tially performed, which helps reduce noise from external sources.
Using nephelometric turbidity units (NTU) standards, we obtain
detectable measurements in a broad range of NTU from 1.95 to
1000. By implementing the proposed methodology, we proposed
a video-based inexpensive yet reliable turbidimetry instrument.
Index Terms— Bacterial growth monitoring, image analysis,
nephelometry, turbidimetry, vision-based measurement (VBM).
I. I NTRODUCTION
T URBIDITY in liquids originates from the light scattering
of suspended particles, which correlates with the hetero-
genicity of the system. When a liquid sample is irradiated
with an electromagnetic wave, its electrical charge oscillates,
and thus, the accelerated electrical charges emit radiation in all
directions, and the resultant secondary emission is the scattered
light [1]. Therefore, light scattering is an important tool for
particle characterization [2]. It is noteworthy that turbidimetry
can be successfully modeled using sphere approximations with
a good correlation with experiments [3].
Since turbidity is a common phenomenon, its measurement
is useful for many technological applications. For instance,
initial water quality assessment requires it to be optically
transparent since it may be traceable to pathogenicity [4]–[6].
Other turbidimetry applications include quantitative assays to
measure bacterial growth [7], enzymes [8], and sulfate ion
content [9], among others.
Manuscript received July 1, 2019; revised January 21, 2020; accepted
March 18, 2020. Date of publication April 7, 2020; date of current version
September 15, 2020. This work was supported in part by Conacyt under Grant
CB-2016-01/286926 and in part by the U.S. Army CCDC under Contract
W911NF1810428. The Associate Editor coordinating the review process was
Jochen Lang. (Corresponding author: Gabriel Luna-Bárcenas.)
Alejandro J. Gimenez, José Mauricio López-Romero, and Gabriel Luna-
Bárcenas are with the Centro de Investigación y Estudios Avanzados del
Instituto Politécnico Nacional Unidad Querétaro, Santiago de Querétaro
76230, Mexico (e-mail: gabriel.luna@cinvestav.mx).
Francisco Javier Medellín-Rodríguez is with FCA, Universidad Autónoma
de San Luis Potosí, San Luis Potosí 78210, Mexico.
Lydia M. Contreras-Martinez and Isaac C. Sanchez are with the McKetta
Department of Chemical Engineering, The University of Texas at Austin,
Austin, TX 78712 USA.
Francisco Villaseñor-Ortega is with the Tecnologico Nacional de Mexico
en Celaya, Celaya 38010, Mexico.
Color versions of one or more of the figures in this article are available
online at http://ieeexplore.ieee.org.
Digital Object Identifier 10.1109/TIM.2020.2985902
0018-9456 © 2020 IEEE. Personal use is permitted, but republication/redistribution requires IEEE permission.
See https://www.ieee.org/publications/rights/index.html for more information.
It is noteworthy that turbidimetry is a robust optical tech-
nique that has been successfully used for many years; in
this regard, instrumental setups have not substantially changed
over time. Turbidity has been typically measured by optical
absorbance using Beer–Lambert law [10], [11]; here, a spec-
trometer is often used, and the loss of intensity of transmitted
light is correlated with the liquid turbidity [10], [12]. Neph-
elometry is another method to measure turbidity, which is
more sensitive to suspended material at low concentrations;
it is based upon measuring the scattered light from a light
beam passing through a liquid. In nephelometry, a photode-
tector is typically placed at a fixed angle of 90◦ from the
beam incidence, allowing more sensitive measurements as
the signal-to-noise ratio (SNR) is higher than absorbed light
measurement [12], [13]. Other approaches have been proposed
to enhance turbidimetry measurements, such as single-photon
counting [14] and optic fiber setups [15]. Previously, our
group studied the high sensitivity of nephelometric techniques
to detect magnetic nanowires suspended in concentrations
as low as 2 ng/ml [16], [17]. Recent studies have also
addressed the use of digital cameras for turbidity measure-
ments, including wastewater evaluation [18] and sludge sed-
imentation [19]. An extensive review of different approaches
of detecting turbidity using computer vision has been reported
elsewhere [20]; in this regard, some of the methods reviewed
are gray histogram analysis [21] and shape detection [22],
among others. In this article, we propose a new algorithm and
approach that may bring advantages for some specific applica-
tions, including the use of small samples and compact setup.
The proposed algorithm can be carried out using minimal
computing power.
With the advent of vision-based measurements (VBMs),
one can produce reliable measurements for many applications
by simply using a camera [23]–[25]. VBM can outperform
old technologies typically based upon expensive apparatus
as spectrophotometers and integrative optical spheres [26].
VBM takes advantage of the great capability of modern video
cameras that capture large amounts of information in relatively
short times.
Based upon the above, the objective of this article is to
implement a VBM methodology that enables us to build an
inexpensive yet highly sensitive turbidimetry apparatus. Our
approach includes the use of an inexpensive video camera
and other simple accessories to build a VBM apparatus.
The proposed technology could potentially perform equally
or better than expensive spectrophotometers and integrative
optical spheres.

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 GIMENEZ et al.: TURBIDIMETRY BY IMAGE DEGRADATION ANALYSIS 7575

Fig. 3. Scattering integration cone due to the use of a sensing array from a
video camera.

Fig. 1. Representation of image capture using a liquid sample as semitrans-

parent medium.

Fig. 2. Representation of pixel value diagram for image captured for different
transparency media. Fig. 4. Turbidimetry setup.

II. M ETHODOLOGIES AND OBTAINED RESULTS

A. Procedure for Image Analysis in Turbidimetry
Measurements
A common and simple practice for turbidity estimation is to
place a liquid sample in front of a black and white background.
Here, the observer can visually distinguish different degrees
of transparency. Similarly, in this article, we capture an image
using a video camera and then applying a mathematical
algorithm to estimate the turbidity. The video camera targets
the liquid sample contained in a transparent cuvette; behind
the sample, a high contrast pattern is placed, as seen in Fig. 1.
The described setup allows us to capture the image of
the reference pattern, which depends upon the transparency Fig. 5. Instrument imaging capture. The area inside the green square is
of the sample. If the sample is transparent, the image will processed as the clear reference, and the area inside the red square is processed
consist of sharp white and black regions. When a sample is as the semitransparent sample.
opaque, the image will appear as plain gray, which means that
the background image is lost. Therefore, if a semitransparent
sample is measured, the image will consist of gray patterns. standard deviation of pixel values as follows. From Fig. 2,
Digitalized captured pixel values by the camera can be rep- one can observe a transparent image in which pixel values
resented by numerical values. In this regard, 0 is for black drastically change from maximum to zero. On the other
and 255 for white; all other values in between represent gray extreme, an opaque image exhibits no changes in the inten-
colors. A schematic of this principle is shown in Fig. 2. sity. Consequently, a transparent sample will yield a large
Based on the methodology mentioned earlier, we propose to pixel standard deviation value, and an opaque sample will
correlate the sample turbidity to the sharpness of the captured yield a standard deviation tending to zero. A semitransparent
image. The sharpness can be quantified by estimating the sample will yield a standard deviation between both cases.

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 7576 IEEE TRANSACTIONS ON INSTRUMENTATION AND MEASUREMENT, VOL. 69, NO. 10, OCTOBER 2020

Fig. 6. Standard deviation changes of reference (red) and sample (blue). Both turbidity measurements are performed with DI water. Noise compensation
(black) is calculated by dividing the sample signal by the reference signal; this method yields a more stable measurement.

The standard deviation for pixel intensity is calculated using

the following equation:

N
i=1 (x i − x̄)
2
s= (1)
N −1
where N is the number of pixels and x is the pixel intensity
value. The procedure is then applied in a differential fashion,
i.e., the relative turbidity of a sample can be calculated by
subtracting the standard deviation of a reference (sref ). From
that of the sample (ssam ) and normalizing by the reference
(sref ) as follows:
ssam
Relative Turbidity = 1 − . (2)
sref
A relative turbidity = 0 implies that the sample has the same
transparency as the reference; relative turbidity = 1 means an
opaque sample, and relative turbidity < 0 could be obtained Fig. 7. Relative turbidity scale (proposed) ranges from zero (DI water
if the sample is clearer than the reference. reference) to one (totally opaque sample) as a function of NTU standardized
samples using formazine as the reference standard. The red line shows the
mathematical correlation of the obtained data to the Beer-Lambert law.
B. Basic Definitions of the Scattering Measurement Principle
When a sample contains suspended particles, the light is
scattered in all directions; after passing through a sample, the closely related to turbidity and can be defined as the scattering
light intensity I is attenuated following the Beer–Lambert law: function f (). This function returns the scattering intensity
at any given solid angle  as follows:
I = Io exp[−(σa + σs )] (3)

where σa and σs are volumetric coefficients of absorption σs = f ()d (4)
and scattering, respectively. The scattering coefficient σs is =2π(1−Cosθ)

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 GIMENEZ et al.: TURBIDIMETRY BY IMAGE DEGRADATION ANALYSIS 7577

Fig. 8. Turbidity measurements of colloidal suspensions. (a) ZnO (40 μg/ml). (b) NiO (200 μg/ml). Measurements are done by triplicate (black, blue, and
red) to assess repeatability and robustness of the proposed turbidimetry method (Right: relative turbidity. Left: NTU scale).

where  is the solid angle from the scattering cone [ = 2π
(1-Cosθ )]; the scattering cone is formed due to the capacity of
the camera sensing array to collect scattered light in a range
of possible directions, as shown in Fig. 3.
It is noteworthy that our proposed instrument has integrating
properties that exhibit improved sensitivity [26], [27]. Further
mathematical analysis should be done to exploit the capacities
of our instrument fully.
C. Instrumental Setup
A USB color camera, 640 × 480 pixels (based on CMOS
GC0308 sensor), is used for imaging capture; the software
interface for reading and processing the input data is coded
in Microsoft Visual Basic 2015. Samples are placed in 3.5-ml
optical cuvettes, and regular bond paper marked with black
ink is used as a background pattern. Background illumination
is provided by one diffused white LED.
3-D printing was used to fabricate the components, as shown
in Fig. 4; the encasement is printed in black polylactic acid
(PLA) plastic. The video camera is 3 cm away from the
samples, the two cuvettes are 2 mm from each other, and the
illuminated background pattern is 2 mm behind the cuvettes.
Fig. 5 shows captured images for both the reference and
sample patterns. Our software is capable of properly adjust
the position of the patterns in two 14 400 (120 × 120) pixel
images (one for the reference and one for the sample). Color
images are converted to greyscale and are integrated for 15 s
to minimize noise. The standard deviation of each image
is calculated using (1), and the sample relative turbidity is
obtained from (2).
Deionized (DI) water is used as a zero-turbidity sample.
In addition, DI water is used to make a software calibration
to compensate for the small inhomogeneities that exist in the
background pattern.
D. Differential compensation
1) Methodology: To assess the presence of noise, a 200-min
standard deviation measurement is performed. The noise com-
pensation experiment is carried out by triplicate to reveal
the constantly changing nature of the measurement. The test
is performed using DI water in both reference and sample
cuvettes. Differential compensation is applied by dividing the
measurement sample standard deviation signal by the reference
signal, as explained in Section II-A.
2) Results: Typical standard deviation values are shown in
Fig. 6. Here, the standard deviation exhibits fluctuations for
periods up to 200 min. It is noteworthy that both sample
and reference are DI water, and the associated signals from

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 7578 IEEE TRANSACTIONS ON INSTRUMENTATION AND MEASUREMENT, VOL. 69, NO. 10, OCTOBER 2020
the reference and instrument are changing similarly. Thus,
it can be inferred that these fluctuations are traceable to noise
rather than simultaneous turbidity changes in both samples.
Consequently, continuous reference compensation is required.
Fig. 6 also reveals that standard deviation drifts ca. 0.5%.
However, using differential compensation, this drift is reduced
fivefold.
E. Vision-Based Measurements Versus Nephelometric
Turbidity Units Using a Formazine Standard
1) Methodology: To assess the validity and robustness of
our proposed methodology, different solutions were prepared
from a 1000 nephelometric turbidity unit (NTU) formazine
certified reference (Sigma Aldrich, TURB1000). Prepared
samples ranging from 0.97 to 1000 NTU [28] were measured
for 2 min. Collected data help define a relative turbidity scale
as a function of the NTU scale aided by a mathematical fitting
based upon the Beer–Lambert equation [(3)].
2) Results: Fig. 7 shows the correlation of our relative
turbidimetry method as a function of standardized NTU. It is
noteworthy that NTU values as low as 1.95 can be detected and
up to 1000 NTU. The proposed relative turbidimetry scale is
based upon the image loss, which is different from the standard
scale based in the concentration of formazine; the coincidence
of both scales is related to the Beer–Lambert law previously
mentioned in (3), using an exponential fit, it can be found σa
and σs , volumetric coefficients, and using these coefficients,
we could convert our data to the NTU standard.
F. Turbidity Measurement Test
1) Methodology: To test our methodology with technolog-
ically relevant particles, we now study the stability over time
of the following colloidal suspensions(∗).
1) Suspension of ZnO powder (Sigma Aldrich 205532,
< 5 μm) in DI water, 40 μg/ml.
2) Suspension of NiO powder (Sigma Aldrich 399523,
325 mesh) in DI water, 200 μg/ml.
(∗) Reference cuvette is filled with DI water, 60-min measure-
ments.
Before turbidity measurements, suspensions were sonicated
for 10 min using a 50-W ultrasonic bath to ensure good
particle dispersion. Experiments were done by triplicate to
assess the repeatability and robustness. Results are converted
to NTU units using the Beer–Lambert equation, as explained
in Section II-E.
2) Results: Aqueous suspensions of ZnO and NiO are tested
for turbidity changes as a function of time, and the results are
shown in Fig. 8.
The behavior, as shown in Fig. 8, reveals the consistency of
our measurements. In this regard, the observed turbidity decay
can be traced to agglomeration driven by flocculation, which
is typically observed in such metal oxide colloids [29].
III. C ONCLUSION
We propose an inexpensive and robust methodology to
measure the turbidity of liquid samples. Our method is based
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upon a VBM setup that takes advantage of commercially
available cameras. A mathematical algorithm based upon the
Beer–Lambert model has been implemented to fit collected
data; sample turbidity is correlated with video image loss.
The procedure involves differential measurements that reduce
fivefold the SNR. In addition, we proposed a new relative
turbidity scale that correlates well with the NTU by testing cer-
tified formazine references. NTU values as low as 1.95 (highly
transparent samples) can be detected and up to 1000 NTU
(highly opaque samples). To assess the validity of the relative
turbidity scale, the stability of aqueous suspensions of ZnO
and NiO were monitored for up to 60 min.
In summary, our technology may prove useful in high-
resolution, low cost, and simple turbidimetry measurements.
This technology may be used as a powerful tool in colloid
science that is relevant in many technological applications.
ACKNOWLEDGMENT
We thank Reina Araceli Mauricio Sanchez for sample
preparation and technical assistance.
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Alejandro J. Gimenez received the Ph.D. degree
in materials science from the Cinvestav Queretaro,
Real de Juriquilla, Mexico, in 2013.
He held a research position at Texas A&M Uni-
versity, College Station, TX, USA, from 2008 and
2010. He is currently a member of The National
Researchers System (SNI LEVEL I). His cur-
rent research interests include nanowire sensors,
video-based instrumentation, paper-based electronic
devices, and the development of electronic research
instruments and tools.
Francisco Javier Medellín-Rodríguez was the
Director of the Faculty of Chemistry (2008-2016),
where he is currently a Research Professor with
the Faculty of Chemistry, Autonomous University
of San Luis Potosí, San Luis Potosí, Mexico. He is
also the Head of the polymers and X-rays areas at
Facultad de Ciencias Quimicas (FCA). He has been
a member of the National Research System since
the year 2000, currently Level II. He is an expert in
polymer physics and chemistry.
Dr. Medellín-Rodríguez was a recipient of the
Universidad Autónoma de San Luis Potosi (UASLP) Prize 2019 (consolidated
researcher).
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7579
Lydia M. Contreras-Martinez received the B.S.E.
degree (cum laude) in chemical engineering from
Princeton University, Princeton, NJ, USA, in 2003,
and the Ph.D. degree in chemical engineering from
Cornell University, in 2008, with a focus on engi-
neering bacterial cells for improved production of
therapeutic proteins.
She was a Post-Doctoral Associate with the
Wadsworth Center, New York State Department of
Health, Albany, NY, USA, where she was involved
in understanding mechanisms of infection in patho-
genic bacteria. She is currently an Associate Professor and Laurence
E. McMakin, Jr., Centennial Fellow of chemical engineering with The Uni-
versity of Texas at Austin, Austin, TX, USA, where she is also a member
of the Institute of Cell and Molecular Biology. She teaches introduction to
chemical engineering computing, thermodynamics, introduction to chemical
engineering analysis, and fundamentals and applications of cellular regulation.
José Mauricio López-Romero received the Ph.D.
degree in physics from the Centro de Investigacioìn y
Estudios Avanzados del IPN, Cinvestav-IPN, Mexico
City, Mexico, in 1993.
He joined as the Chief of the Time and Fre-
quency Division, Centro Nacional de Metrologia,
CENAM, Municipio del Marques, Mexico, in 1994.
In 2015, he joined CINVESTAV Queretaro, Real de
Juriquilla, Mexico, as the Director. He is actively
involved in the development of the Sistema Inter-
americano de Metrología (SIM) time and frequency
comparison network and the SIM timescale. He has served as the Chairman
of the SIM Time and Frequency Working Group Queretaro, Mexico. His
current research interests include time scales generation, primary frequency
standards, cesium fountain clocks, quantum gravimeters, optical frequencies,
and quantum cryptography.
Isaac C. Sanchez currently holds the William
J. Murray, Jr., Endowed Chair Emeritus in engineer-
ing with The University of Texas at Austin, Austin,
TX, USA. In 2019, he retired after serving the
Chemical Engineering Department, The University
of Texas at Austin, for 31 years. In 1996, he was
elected the U.S. National Academy of Engineering.
Francisco Villaseñor-Ortega received the Ph.D.
degree in biotechnology from UAM-Iztapalapa,
Mexico City, Mexico, in 2004.
He is currently a Professor Titular C with the
Department of Biochemical Engineering, National
Technology of Mexico, Celaya, Mexico. He is also a
member of the National Researchers System of Mex-
ico and PRODEP accreditation. He is an Accred-
ited Scientist of the Japan International Coopera-
tion Agency (JICA), Tokyo, Japan, for South–South
Mexico Japan cooperation. His research focuses on
bioprocess engineering, production, and purification of heterologous proteins
synthesized in bioreactors, enzymatic catalysis, and biopolymers.
Gabriel Luna-Bárcenas received the Ph.D. degree
in chemical engineering from The University of
Texas at Austin, Austin, TX, USA, in 1997.
He was a Post-Doctoral Researcher with Princeton
University, Princeton, NJ, USA. He has supervised
20 Ph.D. theses at Cinvestav Queretaro, Real de
Juriquilla, Mexico. He also holds two international
patents. He is heavily involved in industry-related
projects. He received a Fulbright-Garcia Robles
Fellowship from the United States Department of
State, Washington, DC, USA, and holds a National
Researcher System of Mexico Accreditation Level III. He has authored or
coauthored more than 180 scientific articles related to biopolymer synthesis
and characterization for tissue engineering applications.