Autophagy analysis was conducted as the previous description.46 Briefly, GFP‐LC3 plasmid was used to transfect H1299 cells and then treated with trans‐gnetin H and resveratrol for 6 h. Paraformaldehyde (4%) was used to fix the cells and maintained for 15 min. The permeabilized of fixed cells was performed using 0.5% Triton X‐100 in PBS for another 15 min. Sequentially, it was blocked at room temperature using 1% BSA for 1 h and stained with DAPI. Finally, the GFP‐LC3‐containing puncta were measured using laser a scanning confocal microscopy (Leica, Germany).
Laser a scanning confocal microscopy
Manufactured by Leica
Sourced in Germany
Laser scanning confocal microscopy is an imaging technique that uses a focused laser beam to scan a sample and generate high-resolution, three-dimensional images. The core function of this equipment is to provide detailed, non-invasive optical sectioning of specimens, enabling the visualization of specific structures within a sample.
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Lab products found in correlation
2 protocols using laser a scanning confocal microscopy
1
Autophagy Analysis of H1299 Cells
2
Autophagy Analysis of FLCN Mutants
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Autophagy analysis was conducted as the previous description.[27 ] Briefly, GFP‐LC3 plasmid was used to transfect FLCN WT, FLCN S62A, and FLCN S62D HCT116 cells. Paraformaldehyde (4%) was used to fix the cells and maintained for 15 min. The permeabilized of fixed cells was performed using 0.5% Triton X‐100 in PBS for another 15 min. Sequentially, it was blocked at room temperature using 1% BSA for 1 h and stained with DAPI. Finally, the GFP‐LC3‐containing puncta were measured using laser a scanning confocal microscopy (Leica, Germany).
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