Fluorescence laser scanning confocal microscopy

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Sourced in Germany

Fluorescence laser scanning confocal microscopy is an imaging technique that uses a laser to scan a sample and capture high-resolution images by detecting the fluorescence emitted from the sample. The core function of this equipment is to provide detailed, three-dimensional images of biological specimens.

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Fluorescence microscopy (464 citations) Confocal microscopy (351 citations) Confocal laser scanning microscopy (253 citations) Laser confocal microscopy (64 citations) Confocal fluorescence microscopy (40 citations) Laser confocal fluorescence microscopy (6 citations) Scanning laser microscopy (2 citations)

2 protocols using fluorescence laser scanning confocal microscopy

Measuring Mitochondrial Membrane Potential

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The fluorescent probe, JC-1 (Beyotime Biotech, Nanjing, China), was used to measure the MMP according to the manufacturer's instructions, as described in our recent study [38] (link).
The cells were incubated with JC-1 staining solution (5 μg/mL) for 20 min at 28 °C. Then cells were rinsed twice with JC-1 staining buffer and resuspended in 300 μl JC-1 staining buffer. Two laser lines (488 nm and 552 nm) were used to collect fluorescence images (green and red fluorescence) by fluorescence laser scanning confocal microscopy (Leica, Wetzlar, German). The MMP was calculated as the fluorescence ratio of green to red.

Song Y.F., Hogstrand C., Ling S.C., Chen G.H, & Luo Z. (2020). Creb-Pgc1α pathway modulates the interaction between lipid droplets and mitochondria and influences high fat diet-induced changes of lipid metabolism in the liver and isolated hepatocytes of yellow catfish. The Journal of nutritional biochemistry, 80.

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Mitochondrial Membrane Potential Measurement

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Measurements of MMP were made by staining hepatocytes with the lipophilic substance 5,5,6,6-tetrachloro-1,1,3,3-tetraethylbenzimidazolyl carbocyanine iodide (JC-1) according to the manufacturer’s instructions, as described in our recent study [32 (link)]. This dye differentially labels mitochondria according to their membrane potential, emitting in the high orange wavelength for high MMP and in the green wavelength for low MMP. Briefly, the cells were incubated with JC-1 staining solution (5 μg/mL) for 20 min at 28 °C. Then, the cells were rinsed twice with JC-1 staining buffer and resuspended in 300 μL of JC-1 staining buffer. Two laser lines (488 nm and 552 nm) were used to collect fluorescence images (green and red fluorescence) through fluorescence laser scanning confocal microscopy (Leica, Wetzlar, Germany). The MMP was calculated as the fluorescence ratio of green to red.

Song Y.F., Zheng H., Luo Z., Hogstrand C., Bai Z.Y, & Wei X.L. (2022). Dietary Choline Alleviates High-Fat Diet-Induced Hepatic Lipid Dysregulation via UPRmt Modulated by SIRT3-Mediated mtHSP70 Deacetylation. International Journal of Molecular Sciences, 23(8), 4204.

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