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1 Federal Research Center for Virology and Microbiology,

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

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1 Federal Research Center for Virology and Microbiology,

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  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 1,2,31Laboratory 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 Saratov3Department for Microbiology, Biotechnology and Chemistry, Saratov State Agrarian University………………………………………………………………………….4Department for Medical Physics, Saratov State University5Department 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, 410012, Saratov, Russia 5Saratov State Medical University, 410028, 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. 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. 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 # 17-16-01099

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