<title>Real-time monitoring of immunology reactions using computer-aided phase microscope</title>

Abstract

The main advantages of the computer-aided phase microscope (CPM) Airyscan which permits to observe the cell surface and internal structure consists in superresolution and possibility of the dynamic processes registering. The iminunocompetent cells in process ofits activation with different agents were Studied. Several criterions for evaluation of the functional status of immunocompetent cells based on the shape changes were suggested on the basis real-time immunology reaction monitoring. The perspectives ofthe proposed method for immunology testing are discussed. INTRODUCTIONThe recent investigations have shown that there is a substantial difference between phase and amplitude images. So, the new approach to the problem of superresolution based on the features of phase images was proposed in papers1'2. The properties ofhigh order Gaussian beams and wavefront dislocations were at the first time discussed by J.F. Nye and M.V. &rry and later observed experimentally in the microwave patterns by J.V. Hainae .A factor-of-ten over classical diffraction limit enhancement was achieved with a CPM Airyscan. This device also permits dynamic processes registrating at any selected point ofthe phase image.Living cell's investigation and its reaction on external factors represents significant interest for practical medicine and biology.The main aim of this report is the application of CPM Airyscan for real-time monitoring of immunological reactions in human living blood cells. METHODSThe CPM Airyscan is based on optical scheme of well-known Linnik interferometer including reference mirror on piezo-actuator replaceable lens, He-Ne laser (? = 633 urn) and dissector as coordinate-sensitive photodetector. The electronic unit provides processing of the interference signal, dissector scan control and data exchange with IBM PC 386. The software enables pseudocolor display of micrology in the units of equivalent height, 3D-imaging, fragmentation crossections, histograms. The optical magnification is about iø6 times, dimensions of the patterns on the screen may by varied in the internal 64-256 pixels, and the smallest pixel size is about 5 nm for 100A).9 lens. The dynamic processes recording is possible at any selected point of the image with time resolution of 1 ms and Fourier spectra presentation. The mean square fluctuations are 0.5-1.0 urn. Sample preparation. Sample preparation for CPM is very simple. The specimen with cover glass is placed on the mirror reflecting surface which is fixed onto the translation stages of Airyscan. 348 / SPIE Vol. 2329 O-8194-1662-2/95/$6.OO Downloaded From: http://proceedings.spiedigitallibrary.org/ on 06/27/2016 Terms of Use: http://spiedigitallibrary.org/ss/TermsOfUse.aspx