title: Interference reflection microscopy. A quantitative theory for image interpretation and its application to cell-substratum separation measurement.
abstract: We propose a quantitative theory of microscope interferometry where the specimen is illuminated by a cone of monochromatic light of solid angle 0 - 100 degree, corresponding to an illuminating numerical aperture of 0 to approximately 1.2. Computed results compare favorably with photometric measurements of fringe irradiance for a water wedge 0 - 2,000-nm thick. The interpretation of cell-substratum interference images is discussed in relation to the theory. We conclude that in assessing cell-glass separation, the cytoplasmic thickness does in general contribute significantly to the final image, but this contribution is minimized at high illuminating apertures. In these circumstances, however, normal incidence theory is inapplicable and the theory for finite illuminating aperture is essential. Neglect of this fact can lead to errors of up to 100% in estimated cell-glass separation.
mesh_terms: D004247:DNA / Q000032:analysis; D008027:Light; D008433:Mathematics; D008853:Microscopy / Q000379:methods*; D013053:Spectrophotometry / Q000379:methods*
chemical_list: D004247:DNA
references.citation: Nature. 1977 May 12;267(5607):124-8
references.pmid: 16073410
references.citation: J Cell Biol. 1964 Feb;20:199-215
references.pmid: 14126869
references.citation: J Cell Sci. 1978 Feb;29:197-212
references.pmid: 564353
references.citation: Nature. 1977 Aug 25;268(5622):767-9
references.pmid: 895882
references.citation: J Cell Biol. 1966 Sep;30(3):601-21
references.pmid: 5971008
references.citation: Science. 1976 Jan 30;191(4225):399-400
references.pmid: 1246623
references.citation: Exp Cell Res. 1975 Apr;92(1):57-62
references.pmid: 1169157
references.citation: J Mol Biol. 1969 Feb 28;40(1):19-32
references.pmid: 5363346
references.citation: J Cell Sci. 1976 Jun;21(1):129-59
references.pmid: 932106