1. Application of laser scanning speckle-microscopy for high-resolution express diagnostics of chlamydial infection  

2. Irina Subbotina2,3, Nadezhda Filonova2,3, Sergey Zaitsev2, Yury Saltykov2, Tatiana Polyanina2, Anna Lyapina2, Yulia Moiseeva5, Onega Ulianova2, Sergey Ulyanov1,2,4, Irina Kalduzova4, Olga Larionova1,3, Sergey Utz5, Valentina Feodorova 1,2,3 1Laboratory for Molecular Biology of Chlamydia, Federal Research Center for Virology and Microbiology…………………………………………………………………. 2Laboratory for Molecular Biology and NanoBiotechnology, Federal Research Center for Virology and Microbiology, Branch in Saratov 3Department for Microbiology, Biotechnology and Chemistry, Saratov State Agrarian University…………………………………………………………………………. 4Department for Medical Physics, Saratov State University 5Department of Dermatology and Venerology , Saratov State Medical University  

3. 1 Federal Research Center for Virology and Microbiology,
601120, Pokrov, Vladimir region, Russia
2 Federal Research Center for Virology and Microbiology,
Branch in Saratov, Ap. 9, Proviantskaya Street, Box 1580,
410028, Saratov, Russia
3 Saratov State Agrarian University, 1, Theatralnaya Square,
410012, Saratov, Russia
4 Saratov State University, 83 Astrakhanskaya Street,
5 Saratov State Medical University, , Saratov, Russia

4. Method of speckle-microscopy has been adapted to the problem of detection of Chlamydia trachomatis microbial cells. Prototype of laser scanning speckle-microscope has been designed. Spatial resolution and output characteristics of this microscope have been analyzed for the case of scanning of C. trachomatis bacteria inclusions inside the epithelial cells, fixed on the glass. It has been demonstrated, that presence of C. trachomatis microbial cells in the sample can be easily detected using speckle microscopy

5. Optical scheme of speckle-microscope
Chlamydia trachomatis
cell inside the epithelial cell
Speckles
Microobjective
90x (LOMO)
Photodetector
PDA10, (Thorlabs),
connected
to DAC, (NI)
He-Ne laser beam
LGN 205
Microscope slide with
a fixed bacterial cells
Motorized linear stage
(Standa)

6. Amplitude characteristics of scanning speckle-microscope
The amplitude characteristic is close to the linear
one in the range of 0< A/λ <0.075.
A is an amplitude of phase irregularity,
introduced by inhomogeneities of the sample.
If parameter of A/λ > 0.2, then the amplitude
characteristic begins to oscillate in an intricate way.

7. with blood serum of the patient with chlamydial infection.
McCoy cells incubated
with blood serum of the patient with chlamydial infection.
<- Control. McCoy cells
<- 2 hours of incubation
<- 48 hours of incubation
blood serum of the patient contains
Chlamydia trachomatis bacteria

8. Samples for the scanning
Control.
Pure McCoy cells.
McCoy cells do
not contain
Chlamydia
Trachomatis
cells inside.
McCoy cells.
Dyeing by
Romanowsky-Giemsa
McCoy cells.
No dyeing

9. Chlamydia trachomatis microbial cells extracted
Clinical sample.
Chlamydia trachomatis microbial cells extracted
from urethra of the patient
Chlamydia trachomatis cells
fixed on the microspore slide.
Bacteriological smear.
Dyeing by Romanowsky-Giemsa.

10. Scanning of the sample, containing Chlamydia trachomatis cell
Peak corresponds to the moment
of scanning of bacterial cell.

11. Conclusion
It has been proven the high effectiveness of laser scanning speckle-microscopy for high-resolution express diagnostics of chlamydial infection

12. Acknowledgments
This research has been supported by Russian Scientific Foundation, grant #