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Application of frustrated total reflection for studying cells in vitro
Author(s) -
Akatov V. S.
Publication year - 1985
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.1985.tb02569.x
Subject(s) - refractive index , flattening , total internal reflection , optics , light scattering , light beam , scattering , materials science , angle of incidence (optics) , beam (structure) , physics , composite material
SUMMARY A mathematical analysis of the dependence of the light flux scattered upon frustrated total reflection (FTR) on the angle of incidence of the light beam is presented for monolayers of spread and spherical cells. It is shown that using these relationships it is possible to estimate the refractive index of hyaloplasm at cell‐substrate contacts and to study the flattening of the cell surface facing the glass. Analysis of FTR for single cells is presented. A condition is given that must be fulfilled for the brightly shining regions of the cell to be interpreted as the sites of attachment of the cell surface to glass. The condition defines the range of angles of incidence at which FTR does not depend on the optical properties of the cytoplasm portion being at a distance from the glass. The fulfillment of this condition is necessary for spectroscopic and ellipsometric studies of cell surface contacting the glass, by means of FTR. The conclusions are confirmed by experimental data. A peculiarity of light scattering by cells, associated with the coherence of the incident light beam is discussed.