Podocytes were seeded onto type I collagen 4-well culture slides (BD Biosciences, San Jose, CA), cultured for 24–72 hours and stained with AF488 labelled phalloidin. Additional details are provided in Supporting Information .
Type 1 collagen 4 well culture slides
Manufactured by BD
Type I collagen 4-well culture slides are a laboratory equipment product designed for cell culture applications. The slides feature a surface coated with Type I collagen, a common extracellular matrix protein. This product provides a standardized substrate for the in vitro cultivation of cells that require or benefit from a collagen-rich environment.
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2 protocols using type 1 collagen 4 well culture slides
1
Podocyte Adhesion and Cytoskeleton
2
Visualizing Podocyte Actin Cytoskeleton
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Podocytes were seeded onto type I collagen 4-well culture slides (BD Biosciences) and cultured for 24–72 hours. The cells were then washed once with PBS and fixed for 20 minutes with 4% paraformaldehyde. They were permeabilized for 5 minutes with 0.1% Triton X-100 in PBS, followed by blocking for 30 minutes with PBS containing 2% BSA; they were stained for 30 minutes at room temperature with AF488-labeled phalloidin (A12379, Thermo Fisher Scientific). After washing 3 times with PBS, Hoechst 33258 staining was done; then, the slide was sealed with nail polish. Stained specimens were analyzed with a Nikon Eclipse Ti confocal microscope. Images were analyzed with EZ-C1 v.3.7 software, and fluorescence intensities of actin fibers were measured by ImageJ software.
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