Backscattered light confocal imaging of intracellular MTT-formazan crystals

Microscopy Research and Technique
Tytus Bernas, Jurek Dobrucki

Abstract

Metabolically active animal and plant cells reduce MTT tetrazolium salt to a corresponding nonfluorescent formazan. Reduction of MTT by viable cells is exploited in a number of tests widely used in biological research. The aim of this study was to optimize a microscopy method of detecting small crystals of MTT-formazan formed in intact cells maintained in in vitro cultures. We examined scattering properties of small intracellular crystals of MTT formazan and found that the efficiency of light scattering was dependent on wavelength. Small (<3 microm) crystals of MTT-formazan, formed in viable cells, scattered red, but not blue, light. Large crystals, which are formed later at a stage when cells begin to lose viability, scattered both red and blue light. We conclude that optimal detection of early stages of crystallization of MTT-formazan in living cells is possible using confocal microscopy of red, but not blue, scattered light. High contrast and resolution of images can be achieved by filtering out interference effects in the frequency domain.

References

Sep 1, 1979·The Journal of Cell Biology·J Bereiter-HahnB Thorell
May 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·J C SáezD C Spray
Jan 1, 1966·Proceedings of the National Academy of Sciences of the United States of America·B Rotman, B W Papermaster
Jun 9, 1998·Journal of Immunological Methods·A R Mire-Sluis, R Thorpe
Oct 28, 1998·Proceedings of the National Academy of Sciences of the United States of America·Y LiuD Schubert
Jul 20, 2000·Archives of Biochemistry and Biophysics·T Bernas, J W Dobrucki
Nov 4, 2000·Methods in Cell Biology·S L Voytik-HarbinJ P Robinson

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Citations

Mar 10, 2012·Toxicology in Vitro : an International Journal Published in Association with BIBRA·Lanhai LüJie Xu
May 16, 2017·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Joanna SkibaKonrad Kowalski

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