Interference Microscopy in Cell Biophysics. 1. Principles and Methodological Aspects of Coherent Phase Microscopy

Abstract

The purpose of our tandem publications is to review the applications of the coherent phase microscopy to cell biophysics. In this article, we briefly consider the fundamentals and methodological aspects of the coherent phase microscopy (CPM). One of important advantages of this method is a high sensitivity of CPM images to changes in physical-chemical properties of biological samples. The optical path length difference (OPLD), measured with CPM instruments in different domains of an object, serves as the informative optical parameter used for visualization of individual cells or intracellular organelles. Metabolically dependent variations of CPM images reflect changes in the functional state of biological objects. In the next article, we review results of CPM studies of different cells and intracellular energy-transducing organelles.

Quantitative phase imaging of living cells: application of the phase volume and area functions to the analysis of “nucleolar stress”

“…The microscope and the principle of CPM have been discussed elsewhere. [9][10][11][12] One can hypothesize that findings described below can be obtained with phase microscopes other than Airyscan.…”

“…[1][2][3][4][5][6][7][8][9][10][11][12] The phase imaging provides unique opportunity to dissect the biological object using normalized values of optical path difference (OPD). Measurement of this parameter, as well as refractivity, with exceptionally high accuracy allowed for registering the phase thickness (h) and volume-averaged refractive index of erythrocytes, [3][4][5][6][7][8] local fluctuations of cell dynamics, 1,2 and functional responses to extracellular stimuli.…”

“…Measurement of this parameter, as well as refractivity, with exceptionally high accuracy allowed for registering the phase thickness (h) and volume-averaged refractive index of erythrocytes, [3][4][5][6][7][8] local fluctuations of cell dynamics, 1,2 and functional responses to extracellular stimuli. [3][4][5]7,[9][10][11][12][13][14] Still, the literature data on physical parameters of individual cellular compartments remain scarce (see Ref. 15 as an early study).…”

“…The coherent phase microscopy (CPM) [9][10][11][12] has been proved to be informative for quantitative real-time monitoring of a variety of processes in living pro-and eukaryotic cells. 9,[12][13][14] Using this method and a simplified spherical model of a T-lymphocyte, we reported the numerical values of key physical parameters of the nucleus and organelles.…”

Application of Phase Images for Estimation of Peritoneal Macrophages State

“…(Barer, 1957;Byrne et al, 2008;Dunn, 1998;Marquet et al, 2015;Tychinsky & Tikhonov, 2010;Vishnyakov & Levin, 1998)). Interference microscopy and some noninterferometric methods (e.g.…”