Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4462

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Microscopic Analysis of Mitochondrial Transmembrane Potential ({Delta}{Psi}m)

Shailaja Kasibhatla, Gustavo P. Amarante-Mendes, Deborah Finucane, Thomas Brunner, Ella Bossy-Wetzel, and Douglas R. Green

This protocol was adapted from "Apoptosis Assays," Chapter 15, in Cells (eds. Spector et al.). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 1998. This three-volume set is now out of print; however, some of the microscopy methods were republished in Basic Methods in Microscopy, by David L. Spector and Robert D. Goldman.

The first 15% of the full text of this article appears below.


INTRODUCTION

This protocol describes a method for the evaluation of mitochondrial function using the fluorochrome CMXRos. CMXRos is sequestered by actively respiring mitochondria, but washed out when the mitochondrial membrane potential is lost. This analysis can be combined with the TUNEL technique or immunocytochemistry. (Please see Analysis of TUNEL Staining by Flow Cytometry to Detect Apoptosis and TUNEL Staining of Adherent Cells to Detect Apoptosis.) A method for microscopic analysis of CMXRos-treated cells is presented here.


MATERIALS

Reagents

Aqueous fluorescent mounting . . . [Full Text of this Article]


METHOD


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TUNEL Staining of Adherent Cells to Detect Apoptosis
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CSH Protocols 2006: 4433. [Extract] [Full Text]

Analysis of TUNEL Staining by Flow Cytometry to Detect Apoptosis
Shailaja Kasibhatla, Gustavo P. Amarante-Mendes, Deborah Finucane, Thomas Brunner, Ella Bossy-Wetzel, and Douglas R. Green
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