Measurements of mitochondrial NADH pool and NADH production rate in acute brain slices and primary cell cultures using live cell imaging

Angelina I. Dolgikh; Olga A. Stelmashchuk; Andrey Y. Vinokurov; Evgeny A. Zherebtsov; Andrey Y. Abramov

doi:10.1117/12.2590804

4 May 2021 Measurements of mitochondrial NADH pool and NADH production rate in acute brain slices and primary cell cultures using live cell imaging

Angelina I. Dolgikh, Olga A. Stelmashchuk, Andrey Y. Vinokurov, Evgeny A. Zherebtsov, Andrey Y. Abramov

Author Affiliations +

Proceedings Volume 11845, Saratov Fall Meeting 2020: Optical and Nanotechnologies for Biology and Medicine; 118450L (2021) https://doi.org/10.1117/12.2590804
Event: Saratov Fall Meeting 2020, 2020, Saratov, Russian Federation

Abstract

The paper presents results of the estimation of bioenergetic parameters in acute slices of brain rat by measuring mitochondrial pool and rate of NADH production using fluorescence imaging. NAD+ and NADH play crucial roles in mitochondrial energy metabolism due to their participation in the tricarboxylic acid cycle and in electron transport chain. That is why it is essential to know variability of its content in cells. In the study we used an approach based on the registration of auto-fluorescence of NADH. An experimental setup was designed and assembled for the imaging of the acute brain slices but also capable of measuring brain cell cultures. BDL-SMN-375 laser (Becker&Hickl, Germany) was used to excite fluorescence at the wavelength of 375 nm. The parameters of NADH content in both acute brain slices and cell cultures of midbrain and cortex were estimated. The obtained results can be of interest for better understanding of development of neurodegenerative diseases of mitochondrial disfunction origin.

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Angelina I. Dolgikh, Olga A. Stelmashchuk, Andrey Y. Vinokurov, Evgeny A. Zherebtsov, and Andrey Y. Abramov "Measurements of mitochondrial NADH pool and NADH production rate in acute brain slices and primary cell cultures using live cell imaging", Proc. SPIE 11845, Saratov Fall Meeting 2020: Optical and Nanotechnologies for Biology and Medicine, 118450L (4 May 2021); https://doi.org/10.1117/12.2590804

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