These procedures were performed as previously described [14 (link), 28 (link)]. Cells were plated on poly-D-lysine-coated (0.05% w/v; Sigma) 8-well glass culture slides (BD Biosciences) and cultured (48 h) in RPMI 1640 (growth medium) prior to CDDP treatment. For immunostaining, cells were fixed in paraformaldehyde (4%, 1 h, RT), washed in PBS, and blocked with 1% BSA. Mitochondria were visualized by immunofluorescence microscopy, using a mouse monoclonal antibody anti-human Tom 20 (1:100; Santa Cruz Biotechnology) and Alexa Fluor 488 goat anti-mouse secondary antibody (1:500; Invitrogen). Confocal images were obtained (× 100 objective) on an Olympus IX81 inverted microscope with appropriate argon lasers (488 nm). Mitochondrial phenotype of each cell was categorized as being tubular, intermediate or fragmented, as previously described [14 (link)]. At least 100 cells were analyzed per treatment group.
8 well glass culture slides
Manufactured by BD
The 8-well glass culture slides are laboratory equipment designed for cell culture applications. They provide a sturdy, scratch-resistant surface with eight individual wells to accommodate multiple samples or conditions simultaneously.
Automatically generated - may contain errors
2 protocols using 8 well glass culture slides
1
Mitochondrial Dynamics in CDDP-Treated Cells
2
Mitochondrial Morphology Analysis
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These procedures were performed as previously described [13, 26] . Cells were plated on poly-D-lysinecoated (0.05% w/v; Sigma) 8-well glass culture slides (BD Biosciences) and cultured (48 h) in RPMI 1640 (growth medium) prior to CDDP treatment. For immunostaining, cells were xed in paraformaldehyde (4%, 1 h, RT), washed in PBS, and blocked with 1% BSA. Mitochondria were visualized by immuno uorescence microscopy, using a mouse monoclonal antibody anti-human Tom 20 (1:100; Santa Cruz Biotechnology) and Alexa Fluor 488 goat anti-mouse secondary antibody (1:500; Invitrogen). Confocal images were obtained (×100 objective) on an Olympus IX81 inverted microscope with appropriate argon lasers (488 nm). Mitochondrial phenotype of each cell was categorized as being tubular, intermediate or fragmented, as previously described [13] . At least 100 cells were analyzed per treatment group.
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