Designing a Histological Analyzer for Diagnosing Pathomorphological

Changes in Tissues as an Example of Chlamydial Infection

SERGEY KOSTAREV1,2,3, RUSTAM FAYZRAKHMANOV1, NATALIYA TATARNIKOVA2,

OKSANA NOVIKOVA2,3, TATYANA SEREDA2

Perm National Research Polytechnic University,

29, Komsomolski Avenue, 614990, Perm,

RUSSIA

RUSSIA

Abstract: - The article is devoted to the development of a device to study tissue destruction caused by damage

to the histo-hematic barriers of the body, under the influence of chlamydial infection. Cell pathology refers to

changes in its components and ultrastructures with causal relationships. Chlamydiacea is a spectrum of diseases

that, because of their polymorphism, cannot be united by a specific symptom complex, and sometimes affect all

systems and organs. Due to the lack of organotropy and host specificity of the different representatives of

chlamydial structures in cells in electron microscopy). Currently, automation and robotization of research are

penetrating all areas of medicine and veterinary medicine, including histological analysis. Currently, in the

preparation of histological preparation, the technological process is automated in a fragmented way. The

development of a histology robot will help to solve the problem of the shortage of highly qualified histology lab

technicians and pathologists and reduce the burden on medical personnel in general. Processes of automation

and modeling of technological flows and resources in the preparation of histological images and acceptance of

the diagnosis in medicine and veterinary medicine is an urgent tasks. In order to identify pathological processes

at the cellular level, as well as to reduce the error in the performance of histological manipulations, approaches

to the design of a histological robot were developed. The model of the express analyzer structurally consists of

two modules: a histological image preparation module and a pathology recognition module. Laboratory

Received: May 27, 2022. Revised: February 21, 2023. Accepted: March 29, 2023. Published: April 28, 2023.

prokaryotes, but their reproduction is closely

dependent on the host cell. Their unique cycle of

development determines their independent position

in the world of microorganisms, [5], [6]. According

to modern concepts, chlamydiae are small (0.25–1.5

µm in diameter) gram-negative microorganisms that

occupy an intermediate position between bacteria

and viruses. Originally, chlamydiae were classified

as viruses because of their ability to multiply in the

host cell cytoplasm and persist for a long time

vegetative forms, enzymatic activity, sensitivity to

several antibiotics (tetracyclines, macrolides,

quinolones), [7], and the presence of a common

genus-specific antigen. All these facts made it

possible to classify them as bacteria and assign them

to the family Chlamydiaceae, [8]. Chlamydia refers

to such diseases in which the permeability of the

histo-hematic barriers is violated, leading to

degenerative changes in the cellular structures of the

body and, accordingly, to the development of the

lumen of the vessels and are destroyed, which

contributes to the generalization of infection

throughout the body, [10], [11]. At the same time,

we are talking about identifying general patterns of

cell tissue damage. These may include reception of

pathogenic information by the cell and response to

damage, disturbances in cell membrane permeability

and intracellular fluid circulation; cell metabolism

disorders, cell necrosis, cellular dysplasia and

metaplasia, hypertrophy and atrophy, pathology of

development will consist of two modules: a module

for the preparation of histological specimens,

including the necessary procedures for preparing a

slice, fixing the biomaterial, and processing with

reagents, and a module for recognizing morpho-

structural changes in tissues as an example of

pathologies initiated by chlamydial infection.

study of morpho-structural changes in tissues

has received much less attention. Some design

fragments on research automation for the food

industry have been developed, [16], [17].

Technological equipment for individual stages

of histological slice preparation and processing,

[18], [19], [20], is being developed. No

histological analyzer that performs a full cycle

of biomaterial preparation and diagnoses

morpho-structural changes in tissues has been

pH-meter, microtomes (sled, rotary, freezing),

cryostat or cryokite, water bath, table for melting

paraffin sections, set of automatic pipettes,

thermostat, refrigerator, microscope, wiring

machine.

Material for the study was fixed in 10%

formalin. The next day we cut out the slices, and

then we wired them in alcohol of increasing

strength. For histological studies, the material was

fixed in a 4% formaldehyde solution, and the tissues

The approaches of systems analysis theory,

circuit and signal theory, and finite state machines

were used in this work. The Ladder Diagram was

constructed and simulated using CX-One Software.

technique and the pathology indicator technique.

The technological map of the designed device is

shown in Figure 1.

4.2.1 Development of a Fabric Sample

where Load – button to load a tissue sample; Open,

Close – end sensors; Q1 – flag to load biomaterial.

4.2.2 Development of the Pre-Cutting Fabric

Module

The module is designed to cut the required amount

of tissue sample from the incoming biomaterial. The

cutting mechanism performs a reciprocating motion.

flag.

4.2.3 Development of the Module Fixing Material

The module is intended for the fixation of material

in fixing liquid to stop biochemical processes (in

practice different fixers are used: simple ones

containing one component (formalin, alcohol,

acetone) and complex ones containing two or more

components (Carnois liquid: absolute alcohol,

chloroform, glacial acetic acid; Zenker liquid:

where LD3 is the activation of the material fixation

module; Q3 – operation completion flag.

4.2.4 Development of the Technological Break

section. In the pieces not fully fixed on the control

sections, red or pink focal points can be seen. Some

tissues take on a brown color after formalin fixation,

which depends on the transition of oxyhemoglobin

described by equation (4):

fixative, either flowing water or alcohol is used. The

operation of the flushing module is described by

equation (5)

The work of the module will consist of dehydration

of the sample with alcohols of increasing strength.

The module operation will be described by a system

of equations (6)

where RT – is the temperature relay.

4.2.8 Development of a Module for Preparing

Histological Sections

histological sections of 5 μm thickness. This

membrane and chromatin in blue-violet tones. Eosin

stains cytoplasm and some structures (fibers) in

pink-red-orange tones. Figure 2, Figure 3, Figure 4,

and Figure 5 showcase some of the cell

abnormalities studied experimentally in chlamydia

infection and cancer, [14].

For a more in-depth analysis of the cause-and-

effect relationships of pathological processes, we

can consider the sub-tree of pathologies presented in

Figure 9, Figure 10 and Figure 11.

The method of synthesizing a sequential

automaton is to identify the cause-and-effect

patterns that form the tree (Figure 12).

The method of synthesis of the sequential

automaton consists of the construction of the

primary table of states and transitions and further

be coded in the same way (Table 3).

Table 3. Coding of the indicators of the second

The coding of the indicators of the third sublevel

of the "Infectious Specific / Exudation" branch is

shown in Table 4.

sublevel of the "Infectious Specific / Exudation"

branch

The other branches of the pathology tree (Figure

Further, using the method of synthesis of a

sequential automaton, [24], the logical equations for

this branch of pathologies were synthesized:

where c – trigger (Figure 13).

Similarly, we can obtain logical equations for the

other branches of the pathology tree.

The neural network method is currently widely

used for the development of pattern recognition

Currently, no fully automated histological analyzer

implemented in the combinational scheme of the

device. Structurally, the express analyzer consists of

formation. The recognizer block includes

recognition programs based on neural networks and

methods based on the definition of pathology

indicators. Simulation modeling was performed to

determine the indicators of the types of pathologies,

which are represented in the form of a tree structure.

The developed method of determining indicators of

diseases is proposed to be used when designing an

automated histological analyzer. The developed

software-diagnostic complexes will allow the

epidemiological situation. Designing and

[1] Quinn, T.C., Gaydos, C., Shepherd, M., Bobo,

[2] Kochetova, O.V., Tatarnikova, N.A.,

[3] Bommana, S., Polkinghorne, A., Mini

[5] Tood, W.J., Doughri, A.M., Storz, J.,

[6] Stephens, R.S., Kalman, S., Lammel, C., et

[8] Semenov, V. M., Clinical and epidemiological

[9] Tatarnikova, N.A., Kostarev S.N., Sereda,

[12] Nikitaev, V.G., Berdnikovich, E.Yu., Strategy

[14] Kostarev, S.N., Sereda, T.G., Tatarnikova,

[15] Kostarev, S.N., Sereda, T.G., Tatarnikova,

[17] Kostarev, S.N., Novikova-Kochetova, O.V.,

[18] Zelentsova, E.A., Yanshole, V.V.,

[21] Afanasyev, I., Yurina, N.A., Kotovsky, E.F.,

[22] Bugrova, M.L., Blagova, N.V., Timofeeva,

[24] Kostarev, S.N., Tatarnikova, N.A., Novikov

[26] Rotin, D.L., Petrovichev, N.N., Pavlovskaya,

[27] Kochetova, O.V., Sereda, T.G., Development