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Updated: Jul 11, 2025
Autofluorescence Imaging to Evaluate Cellular Metabolism
Published on: November 15, 2021
High levels of FAD autofluorescence indicate pathology preceding cell death
Ekaterina O Bryanskaya1, Andrey Y Vinokurov1, Angelina I Dolgikh1
1Orel State University, 95 Komsomolskaya str, Orel 302026, Russia.
Abstract:
Flavin adenine dinucleotide (FAD) autofluorescence from cells reports on the enzymatic activity which involves FAD as a cofactor. Most of the cellular FAD fluorescence comes from complex II of the electron transport chain in mitochondria and can be assessed with inhibitor analysis. The intensity of FAD autofluorescence is not homogeneous and vary between cells in tissue and in cell culture types. Using primary co-culture of neurons and astrocytes, and human skin fibroblasts we have found that very high FAD autofluorescence is a result of an overactivation of the mitochondrial complex II from ETC and from the activity of monoamine oxidases. Cells with high FAD autofluorescence were mostly intact and were not co-labelled with indicators for necrosis or apoptosis. However, cells with high FAD fluorescence showed activation of apoptosis and necrosis within 24 h after initial measurements. Thus, high level of FAD autofluorescence is an indicator of cell pathology and reveals an upcoming apoptosis and necrosis.
Insights
High flavin adenine dinucleotide (FAD) autofluorescence signals cell pathology. This elevated FAD fluorescence indicates impending apoptosis and necrosis in cells, even if initially appearing intact.
Area of Science:
- Cellular biology
- Biochemistry
- Mitochondrial function
Background:
- Flavin adenine dinucleotide (FAD) autofluorescence reflects cellular enzymatic activity.
- Mitochondrial complex II and monoamine oxidases are primary sources of cellular FAD.
- FAD fluorescence intensity varies significantly between cells.
Purpose of the Study:
- To investigate the implications of high flavin adenine dinucleotide (FAD) autofluorescence in cellular pathology.
- To determine if elevated FAD autofluorescence predicts cell death.
Main Methods:
- Utilized primary co-cultures of neurons and astrocytes, and human skin fibroblasts.
- Assessed FAD autofluorescence intensity and correlated it with cell viability markers.
- Employed inhibitor analysis targeting mitochondrial complex II and monoamine oxidases.
Main Results:
- Very high FAD autofluorescence was linked to overactivation of mitochondrial complex II and monoamine oxidases.
- Cells with high FAD autofluorescence were initially intact, showing no signs of necrosis or apoptosis.
- These high FAD fluorescence cells exhibited subsequent activation of apoptosis and necrosis within 24 hours.
Conclusions:
- Elevated flavin adenine dinucleotide (FAD) autofluorescence serves as a sensitive indicator of cellular pathology.
- High FAD autofluorescence predicts the onset of apoptosis and necrosis in cells.
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