Immunocytochemistry of mitochondrial damage.

B-CLL cells were treated with the indicated drugs for 8 hours. Cells were stained for mitochondria membrane potential with MT orange (red, A) for 30 minutes, pelleted on a slide at 300g, fixed in 4% paraformaldehyde, and stained for cytochrome c (green, B) with monoclonal antibody and secondary antibody Alexa 488. DNA was visualized with DAPI (blue, C). (D) Composite image. Arrows show cells affected by the drug, where cytochrome c release is noticeable. Images were obtained with a Zeiss Axioscope microscope, with CCD camera.