Protocol

Molecular Chaperones

Volume 787 of the series Methods in Molecular Biology pp 231-244

Date:

Determination of Cell Survival or Death

  • Alexander E. KabakovAffiliated withDepartment of Radiation Biochemistry, Medical Radiology Research Center
  • , Vladimir A. KudryavtsevAffiliated withDepartment of Radiation Biochemistry, Medical Radiology Research Center
  • , Vladimir L. GabaiAffiliated withDepartment of Biochemistry, Boston University Medical School Email author 

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Abstract

Cell death (in particular, apoptosis and necrosis) is accompanied by appearance of certain hallmarks that are manifested as specific alterations in cellular membranes, cytoplasm, nucleus and mitochondria. Some of those hallmarks are easily detectable in situ and, therefore, they can be applied for the assessment of dying or dead cells. In turn, there are also signs of viable cells that include a set of features, such as normal functioning of their membranes and organelles, ability to proliferate, etc. The present chapter provides descriptions of several convenient methods for quantitative determination of dead (apoptotic and necrotic) cells and also methods for determination of survived and viable cells. Here, we describe in details the methods of annexin V/propidium iodide (PI) staining, TUNEL assay, Hoechst/PI staining, MTS tetrazolium assay, and colony formation assay, with the principles, advantages, and drawbacks of each technique.

Key words

Apoptosis Necrosis Annexin V TUNEL Colony formation assay Tetrazolium assay