12 February 1999 Real-time monitoring of the functional status of platelets treated by Infukoll using a computer-aided laser phase microscope
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Combined analysis of optic-geometrical characteristics makes it possible to comprehensively evaluate the morphological and functional state of the cytological object, which can not be done during visual observation. The technique is discussed for real-time monitoring of the functional status of platelets using computer-aided phase microscope (CPM) 'Cytoscan'. High accuracy and sensitivity of CPM with respect to determination of local temporal phase make it possible to register the dynamic processes in the voluntarily chosen points and sections of micro-objects, to obtain the Fourier's spectra and other characteristics suitable for statistical analysis. Human platelets were prepared from venous blood of healthy donors and pregnant women by standard methods, suspended in culture medium 199 and treated by different doses of 6% Infukoll HES. Nonfixating and nonstaining cells were studied with CPM: height accuracy 0.5 nm, magnification 1000, acquisition time 4 - 30 s. In our experiments we used time resolution about 0.03 s and 30 x lens with numerical aperture 0.65. During investigations of temporal processes a certain section was chosen in the topogram of cell image and local values for the phase of scattered wave in each of the points of the chosen cell's profile were measured. On the basis of the results of automated phase image analysis of optic-geometrical characteristics of living cells, the new quantitative express-method for evaluating of the functional status of human platelets was developed and tested. The structural changes of cells were visualized in alteration of 3-D images, phase profiles, in the decrease of mean cell phase diameters, heights, volumes, in disturbance of histograms of phase heights distribution by cell image points. New data on the behavior of platelets treated by Infukoll in vitro and in vivo were obtained. Analysis of intracellular dynamics was allowed to characterize the cell's regions of maximal activity, but the intensity of processes taking place in these regions in accordance with frequencies as well. The difference between the calculated values of frequencies (from 0.1 to 17 H) is associated with the influence of Infukoll on the metabolism and functional activity of living platelets.
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Irina A. Vasilenko, Irina A. Vasilenko, Svetlana Babakova, Svetlana Babakova, Elena Kiseleva, Elena Kiseleva, Adyan Dyugeev, Adyan Dyugeev, Alexander A. Konradov, Alexander A. Konradov, Vladimir Shabalin, Vladimir Shabalin, "Real-time monitoring of the functional status of platelets treated by Infukoll using a computer-aided laser phase microscope", Proc. SPIE 3567, Optical and Imaging Techniques for Biomonitoring IV, (12 February 1999); doi: 10.1117/12.339179; https://doi.org/10.1117/12.339179

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