We propose using the method of holographic microscopy to detect fine morphologic changes in living cells. An “LSM 510” laser confocal scanning microscope is modified to allow recording digital microholograms which can be used to reconstruct the amplitude and phase of the radiation transmitting through the sample. Measuring the phase increment of the object beam in cells and the intercellular space yields information on the optical length of the ray path in the cells (spatial dimensions and the refractive index), which in turn contains information on changes in the morphology and intracellular contents. Calcium activity is studied by means of fluorescent microscopy which makes it possible to detect minor variations in the intracellular concentration of calcium ions. By studying the dynamics of calcium oscillations and variations in the optical thickness, conclusions are made about the interrelation of functional and morphological variations, and comparative analysis of these variations is performed.
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Translated from Izvestiya Vysshikh Uchebnykh Zavedenii, Radiofizika, Vol. 57, No. 8–9, pp. 646–653, August–September 2014.
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Murav’eva, M.S., Dudenkova, V.V., I.Rybnikov, A. et al. Parallel Monitoring of Living Cell Cultures by Means of Digital-Holography and Fluorescent Microscopy. Radiophys Quantum El 57, 577–583 (2015). https://doi.org/10.1007/s11141-015-9541-1
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DOI: https://doi.org/10.1007/s11141-015-9541-1