Development of hardware-software microscopy complex for the
study of buccal epithelial cells
A. A. Rumyantseva, G. A. Kolokolnikov, A. V. Samorodov, A. K. Volkov
Bauman Moscow State Technical University, Moscow, Russia
Abstract The paper is devoted to the development of
hardware-software microscopy complex for the study of buccal
epithelial cells. The goal and objectives of the work were
determined, and the approach to achieve them was chosen. A
medical problem was formulated and the biophysical bases for
the study of the electrokinetic properties of buccal epithelial
cells were described. System-wide solutions were proposed in
the form of a developed biotechnical system scheme. A
structural-functional model of the system processes was
developed in accordance with the IDEF0-notation. The
methodology of buccal epithelial cells exploration was proposed.
The hardware implementation and the developed specialized
software used in the research were described. Further
directions of project development were proposed.
Index Terms — buccal epithelium, cells, hardware-software
complex, image processing, microscopy.
I. INTRODUCTION
The relevance of buccal epithelial cells research lies in
cells morphological structure. The cells have an appearance
of polygonal or oval shaped thin plates (Fig. 1), and line the
mucous membrane of the oral cavity. They are playing a
role of the “border” between the internal and external
environment of a body [1, 2].
Fig. 1. Buccal epithelial cells under a microscope
The purpose of the work is to develop hardware-software
complex for studying the relationship between mobility of
the buccal epithelium cells nuclei and human physiological
parameters. The following tasks were completed to achieve
this goal:
− biophysical foundations of buccal epithelial cells nuclei
mobility were studied;
− scheme of biotechnical system was developed;
− structural-functional model of system processes was
developed;
− hardware and software implementation was proposed.
The cells of buccal epithelium reflect the state of the
human body. In particular, they show a response of the
organism to effects that have just occurred (physical,
chemical, pharmacological, etc.).
Myachina et al. have showed relationship between
properties of the buccal epithelium and hypertension,
adverse environmental factors, and Kalaev et al. showed
connection between the buccal cells state and the
inflammation of periodontal soft tissues. [3, 4, 5]. It is also
known that there is a possibility of diagnosing soft tissues
inflammation of periodontal disease at an early stage [6]
with utilization of proposed methodology. Also, study of
buccal epithelial cells allows to assess the overall
physiological state of the human body during physiotherapy
and can serve as an additional parameter for non-invasive
diagnosis of the chronic stress effects [7, 8].
II. MEDICAL TASK
The study of buccal epithelium is based on the method of
intracellular microelectrophoresis, developed by V.G.
Shahbazov and co-authors [9]. This technique allows to
explore electrokinetic parameters of cells.
The biophysical bases of electrokinetic phenomena lie in
such physical processes as flow potential and subsidence
potential. The cause of electrokinetic phenomena is the
presence of a double electric layer at the phase boundary,
which is a conventional flat capacitor, the inner side of
which is charged with ions, and the outer side - with
counterions. In this case, one part of the ions is located near
the phase boundary surface (adsorption), the second part is
in the diffusion part [2].
According to modern concepts, counterions are firmly
bound to a solid surface as a result of both electrostatic and
adsorption forces and form a so-called dense adsorption
layer of counterions. The thickness of the layer is
determined by the size of counterions and is of several
molecular diameters (about 1 nm) [10].
The remaining counterions, which are at a greater distance
from the surface, form a diffuse layer that is held near the
surface only by electrostatic forces. The ions in the dense
part are considered stationary, and the ions in the diffuse
part are mobile. As a result of thermal movement, the
counterions of the diffuse layer can move deep into the
liquid. Effective thickness of diffuse layer is equal to the
distance at which the surface potential decreased by a
factor of an exponent. The diffuse layer thickness depends
on the concentration of electrolytes in a liquid matrix [2].
Decrease of the field potential in a double electric layer
takes place abruptly within the adsorption region and more
smoothly (exponentially) in the diffuse layer region [10].
When the relative movement of phases occurs, the double
electric layer breaks along the slip plane. In this case,
counterions of the adsorption layer do not leave the surface,
and counterions of the diffuse layer move together with the
liquid phase. The potential arising on the slip plane when the
ions of the diffusion layer are detached from the solid phase
is called the electrokinetic, or -potential. The slip plane can
be slightly shifted from the boundary of the adsorption and
diffuse layers, which depends on such factors as the speed of
relative phase shift, the viscosity of the medium.
Accordingly, the value of -potential depends on these
factors. Therefore, the -potential is close to the value of the
diffuse layer potential: • _d. The value of -potential
ranges from a few hundredths to tenths of a volt [2].
The presence of the -potential is manifested in the
electrokinetic phenomena, and the electrokinetic phenomena
underlie the methods for measuring the -potential.
III. DEVELOPMENT OF SYSTEM-WIDE SOLUTIONS
The system being developed belongs to the group of
devices for analyzing electrokinetic and morphological
parameters of biomedical specimens. It is designed to assess
the general physiological state of a patient. For a detailed
description of the system, a diagram of the biotechnical
system, the structural-functional model of the processes and
the methodology for conducting research of the buccal
epithelial micro-specimen are developed. The proposed
solutions are built in accordance with the methodology of
Fig. 2. Scheme of biotechnical system for the study of electrokinetic properties of buccal epithelial cells.
designing automated microscopy systems developed at
BMSTU [11].
A. Development of biotechnical system
The scheme of the biotechnical system for the study of
electrokinetic properties of buccal epithelial cells is
presented in Fig. 2. The prepared patient's cytological
material placed in a specialized chamber for
microelectrophoresis serves as a biological object. The
chamber is represented by a glass slide with a fixed
electrode system. The prepared specimen is located on a
scanning stage of a laboratory microscope.
The physical impact block is represented by a microscope
illumination system and a microelectrophoresis system. At
first, microelectrophoresis system is connected to the
electrode system, then the required mode is set.
The optical system of the microscope together with the
image capture module represent the measuring transducer.
Incident light from the microscope illumination system
passes through the biological object with selective
absorption. Passed light carries information about the object
and falls on the matrix of a digital camera. Registered
images are transferred to a single board computer acting as a
control unit.
Movement automation and focus automation devices are
also used in the automated microscopy complex. To move
biological object motorized scanning stage is used. To
obtain clear images, the focus automation device is used,
which is represented by a stepper motor, a driver, and an
endstop sensor.
An information processing unit of the automated
microscopy complex includes a control module and a
workstation. The workstation monitor and the local interface
module play a role of an information display device. This
device offers graphical user interface for medical staff.
Description of device workflow is presented below. At
the moment of single board computer sending command to
the stepper motors, the stage with the specimen moves along
the X and Y axes to perform scanning. Also, a signal from
the single board computer is sent to the third stepper motor
to control the movements of the specimen along the Z axis.
This ensures focusing.
At specified moments, control signals are sent to the
image capture module. As a result, an image or video of the
specimen is recorded. Then the recorded data is transferred
from the single board computer to the workstation for
further processing.
The presented system “control module - workstation” can
implement the functionality of remote access using the
Internet connection. In this case, the specialist can remotely
analyze the data, meanwhile the preparation is performed by
a laboratory technician working directly with the
microscope.
After data aggregation at the workstation postprocessing
and automated analysis of the recorded data is carried out.
B. Development of structural-functional model system
processes
To describe the information processes occurring in
automated microscopy complex (AMC) for the study of the
electrokinetic properties of buccal epithelial cells, a
structural-functional model of processes was developed
using IDEF0-notation (Fig. 3). A functional set of the AMC
was decomposed. The developed model allows to evaluate
the necessary elements for the operation of the system and to
plan the development of the project. The diagram shows the
input (arrows on the left border of process object) and
Fig. 3. IDEF0-diagram of AMC for the study of the electrokinetic properties of buccal epithelial cells.
output elements (arrows on the right border). Also, the
control actions (above process object) and the elements
required to perform the work (below) are shown in the
diagram.
The main application scenario of the developed AMC is to
study the patient's biological microsample in order to assess
the patient's physiological state. Functional set of AMC is
decomposed into the following processes:
− control AMC;
− set up microelectrophoresis module;
− capture video data;
− analyze video data;
− assess physiological state of a patient.
The control function organizes communication between
all components of the system: the workstation and the
automated microscopy complex. Also, this process includes
the subprocesses of:
− entering personal data of medical staff carrying out the
research in order to gain access to the system;
− choosing operation mode;
− expert-doctor controlling of the research results.
This process should meet the requirements for security of
personal data of both medical personnel and the patient.
The next functional unit is setting up the
microelectrophoresis module. It involves setting the required
parameters of the current, voltage, shape and frequency of
the signal applied to the electrode system. The research of
cells electrokinetic properties can be carried out depending
on the listed parameters. The parameters of the
microelectrophoresis module act as controlling parameters
for the next functional unit and are also used in the patient's
physiological state assessment unit as input parameters.
The functional block of video data registration is
decomposed into:
− placing biological sample into microelectrophoresis
chamber;
− detecting region of the sample with buccal cells;
− choosing sample sections for registration;
− recording video data at the chosen points.
The functional block of video data analysis receives the
registered data and performs the following operations
sequentially:
− detecting cells and nuclei in a video record;
− extracting morphometric parameters of cells and nuclei;
− detecting presence of nuclei fluctuations;
− extracting nuclei fluctuations parameters.
The extracted morphological and kinetic parameters of the
cells are transmitted to the patient assessment unit, along
with the information about parameters of the electrical
signal applied to the electrode system. The unit assesses
correlation between the buccal epithelium cells
characteristics and parameters of the applied electric field.
Based on the obtained data, a medical conclusion is formed,
which is verified by an expert-doctor.
C. Methodology of the buccal epithelial cells research
An order of interaction procedures between medical staff
and patient during the research is shown below:
− taking a biological sample from a patient by scraping
cells from the inside of a cheek;
− sample preparation;
− launching the automated microscopy complex;
Fig. 4. Electrical block diagram of the automated microscopy complex for studying the electrokinetic properties of cells.
− placing microsample into microelectrophoresis chamber
on the scanning stage;
− connecting microelectrophoresis system and setting up
its parameters;
− launching video registration software;
− launching automatic video analysis software;
− automatic assessment of patient physiological state based
on the extracted characteristics of cells and
electrophoresis parameters;
− research results verification by expert-doctor;
− forming a medical opinion.
Depending on the sample preparation method, the quality
of preparation may vary. Therefore, the working area in
which the concentration of cells will be sufficient for
analysis should be located at the first step of research
procedure.
IV. HARDWARE AND SOFTWARE IMPLEMENTATION
A. Hardware
Fig. 4 shows the electrical block diagram of the automated
microscopy complex for studying the electrokinetic
properties of cells.
The main component of the automated microscopy
complex is the Raspberry Pi 3 Model B single-board
computer, which runs the software and establishes
communication with the workstation using Internet
connection. For local interaction with the complex a touch
screen module is intended. Image capturing is carried out
with the help of a camera module connected with a
specialized board. The movement of microsample is carried
out by a motorized scanning stage, which is driven by
stepper motors. Stepper motors and endstop sensors are
connected with a separate board. These boards are located in
the interface control unit (Fig. 5). A separate element of the
system is a microelectrophoresis module with an electrode
system required for studying buccal epithelial cells.
Fig. 5. Internal arrangement of the interface control unit: 1 –
single-board computer; 2 – main connection board; 3 – stepper
motor drivers; 4 – stepper motor connectors; 5 – LED indication
board; 6 – fan of a cooling system.
B. Software
During the development of hardware-software microsopy
complex, the following technological stack was utilized:
− Debian as an operating system;
− Bash for administering software packages;
− Python 3.7 as the main programming language of the
Fig. 6. IDEF0-diagram of video data analysis.
project;
− OpenCV programming package for image processing and
analysis;
− RPi.GPIO for controlling general purpose inputs and
outputs of single-board computer;
− a range of auxiliary packages (NumPy, Matplotlib, etc.).
Algorithm of system workflow is described below.
After launching the ACM, the medical personnel
conducting the research are authorized.
Next, the program sets the parameters of the
microelectrophoresis module. Parameters are saved for
further analysis.
After the microsample is placed under a microscope, the
camera module is launched. Taking into account the
specifics of sample preparation, at the first stage it is
required to determine the working area. To do this, a rapid
scan of the sample is performed. After that step, the working
area with sufficient amount of cells is localized. Video data
is recorded in the regions of interest, which are selected from
the working area. Cells and nuclei of the buccal epithelium
are detected in the recorded video (Fig. 6). It is proposed to
apply the Niblack binarization algorithm [12] for
segmentation of epithelial cells. After segmentation step
morphological parameters are determined for each selected
object by means of a software library Scikit-Image.
Next, the characteristics of fluctuating segmented nuclei
are estimated using object tracking [13] and particle image
velocimetry approaches. The processes of video registration
and assessement of cell nuclei fluctuation parameters are
carried out for several regions of interest. Statistical
processing is applied to the obtained data to evaluate the
microsample parameters in general.
The obtained sets of morphometric and kinetic
characteristics of cells are compared with the parameters of
microelectrophoresis, after which the electrokinetic
properties of the cells are determined, reflecting the general
physiological state of the patient. Upon completion of the
automatic analysis, the results are provided to the expert
doctor for verification. After that a medical conclusion is
formed.
V. CONCLUSION
In the course of the carried out work:
− medical task was formulated and the biophysical
foundations of electrokinetic phenomena of buccal
epithelium cells were determined;
− system-wide solutions were proposed in the form of a
biotechnical system;
− structural-functional model of system processes was
developed;
− methodology for biological microsample research was
proposed;
− hardware of AMC was developed;
− specialized software for biological microsample research
was developed in accordance with the proposed
methodology.
The offered method for studying the electrokinetic
properties of buccal epithelial cells will allow diagnosing a
number of diseases at an early stage, including diabetes
mellitus, hypertensive diseases, diseases of the respiratory
system, assessing the body's response to external forcings,
assessing the quality of a selected therapeutic treatment [3, 4,
8].
Further directions for the development of the project
include finding dependencies between the parameters of
electrical effects, morphological parameters, cell and nucleus
fluctations, and the physiological state of the patient
organism.
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