Interference Microscopy in Cell Biophysics. 2. Visualization of Individual Cells and Energy-Transducing Organelles

Abstract

The coherent phase microscopy (CPM) provides a convenient and non-invasive tool for imaging cells and intracellular organelles. In this article, we consider the applications of the CPM method to imaging different cells and energy-transducing intracellular organelles (mitochondria and chloroplasts). Experimental data presented below demonstrate that the optical path length difference of the object, which is the basic optical parameter measured by the CPM method, can serve as an indicator of metabolic states of different biological objects at cellular and subcellular levels of structural organization.

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.…”

“…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. 13 In this study, we applied the developed algorithms to the analysis of a more complex object, that is, a human colon carcinoma cell (HCT116 cell line) exposed to the antitumor drug actinomycin D (Act D).…”