Effect of Diffraction on the Results of Quantitative Cytophotometry
I. G. Pal'chikovaa1, L. V. Omel'yanchukc2, E. S. Smirnovb3
1 Technological Design Institute of Scientific Instrument Engineering; Novosibirsk State University
2 Institute of Molecular and Cell Biology, Siberian Branch, Russian Academy of Sciences
3 Novosibirsk State University
palchikova@tdisie.nsc.ru
Keywords: Fresnel diffraction, point spread function of the microscope, DNA, cytophotometry
Pages: 92-101
Abstract
The influence of diffraction on the results of computer cytophotometry has been explored through experiments and theoretical analysis. The relative error introduced by diffraction is shown to depend significantly on the optical characteristics of the object: size and absorption. In computer cytophotometry, the choice of standards having a lower DNA concentration in nuclei with large sizes is found to be favorable for measurements of the length of the genomes of various organisms. It is proved theoretically and experimentally that the systematic error due to diffraction can be accounted for without the digital processing of each micro-image involving deconvolution procedures with the point spread function of the microscope.
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