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Coherent phase microscopy in cell biology: visualization of metabolic states
Authors:Vladimir P. Tychinsky  Tatyana V. Vyshenskaya
Affiliation:a Moscow State Institute for Radioengineering, Electronics and Automation, prosp. Vernadskogo 78, 119454 Moscow, Russia
b Faculty of Physics, M.V. Lomonosov Moscow State University, 119992 Moscow, Russia
Abstract:
Visualization of functional properties of individual cells and intracellular organelles still remains an experimental challenge in cell biology. The coherent phase microscopy (CPM) provides a convenient and non-invasive tool for imaging cells and intracellular organelles. In this work, we report results of statistical analysis of CPM images of cyanobacterial cells (Synechocystis sp. PCC 6803) and spores (Bacillus licheniformis). It has been shown that CPM images of cyanobacterial cells and spores are sensitive to variations of their metabolic states. We found a correlation between one of optical parameters of the CPM image (‘phase thicknesses’ Δh) and cell energization. It was demonstrated that the phase thickness Δh decreased after cell treatment with the uncoupler CCCP or inhibitors of electron transport (KCN or DCMU). Statistical analysis of distributions of parameter Δh and cell diameter d demonstrated that a decrease in the phase thickness Δh could not be attributed entirely to a decrease in geometrical sizes of cells. This finding demonstrates that the CPM technique may be a convenient tool for fast and non-invasive diagnosis of metabolic states of individual cells and intracellular organelles.
Keywords:CPM, coherent phase microscopy   DCMU, 3-(3,4-dichlorophenyl)-1,1-dimethylurea   CCCP, carbonylcyanide-3-chlorophenyl hydrazone
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