Quantifying Cardiomyocyte Apoptosis via TUNEL

Visualization of apoptotic cardiomyocyte was achieved in formalin-fixed, paraffin-embedded ventricular sections by the TUNEL technique [15 (link), 24 (link)] using an ApopTag-peroxidase kit (Chemicon International, Inc., San Francisco, CA). Slides were counterstained with methyl green for detection of non-apoptotic nuclei. Enumeration of CM nuclei (both apoptotic and non-apoptotic) was determined within a reference area using an unbiased 2-dimension rule [28 (link)] as described previously [12 (link), 24 (link)]. For each mouse, at least 50 grid fields were counted. The rate of apoptosis was expressed as the percentage of the TUNEL-positive nuclei per total nuclei (apoptotic and non-apoptotic) present within the reference area [12 (link), 24 (link)].

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Hasan K.M., Friedman T.C., Parveen M., Espinoza-Derout J., Bautista F., Razipour M.M., Shao X.M., Jordan M.C., Roos K.P., Mahata S.K, & Sinha-Hikim A.P. (2020). Electronic cigarettes cause alteration in cardiac structure and function in diet-induced obese mice. PLoS ONE, 15(10), e0239671.

Publication 2020

ApopTag-peroxidase kit

Apoptotic Cardiomyocyte Embedded paraffin Formalin Methyl green Mouse Nuclei Peroxidase Tunel Ventricular

Corresponding Organization : University of California, San Diego

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Percentage of TUNEL-positive nuclei per total nuclei (apoptotic and non-apoptotic) within the reference area

control variables

Formalin-fixed, paraffin-embedded ventricular sections
TUNEL technique using an ApopTag-peroxidase kit
Methyl green counterstaining for detection of non-apoptotic nuclei
Enumeration of cardiomyocyte nuclei (both apoptotic and non-apoptotic) within a reference area using an unbiased 2-dimension rule
At least 50 grid fields counted per mouse

controls

None explicitly mentioned

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