PMID: 13563550Jul 25, 1958Paper

The two-wavelength method of microspectrophotometry. III. An extension based on photographic color transparencies

The Journal of Biophysical and Biochemical Cytology
M L MENDELSOHN

Abstract

The two-wavelength method of microspectrophotometry corrects for distributional error and measures the amount of absorbing material by taking advantage of certain spectral characteristics of the specimen. Under certain circumstances, such as the absorption of nucleic acids in the ultraviolet and of black or multiple stains in the visible, the spectral characteristics are not suitable for the application of the method. To circumvent this, a photomicrograph of the object is taken with monochromatic light of a suitable wavelength. A second plate is exposed as a contact print of the photomicrograph and is developed in the presence of a coupling agent. After bleaching and fixation, the positive appears as a monochromatic color transparency. Two-wavelength analysis of such a transparency can be made in terms of the new color. The measurements will be free of distributional error and can be equated to the original object. The necessary formulae are derived, and a method which has proven suitable for color development is given. The photographic and the direct two-wavelength method were found to give equivalent results when both were used on the same series of liver nuclei. The application of the photographic method to ultraviolet absor...Continue Reading

References

Jul 25, 1958·The Journal of Biophysical and Biochemical Cytology·M L MENDELSOHN

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Citations

Nov 1, 1977·The Histochemical Journal·J P Mitchell
Jun 22, 1963·Nature·A KURNATOWSKI, R G WILLIGHAGEN
Sep 29, 2020·Cytometry. Part a : the Journal of the International Society for Analytical Cytology
Jan 1, 1978·Progress in Histochemistry and Cytochemistry·M FukudaS Fujita

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