Publication details

Detection of apoptosis in paraffin embedded tissues: the influence of tissue type and fixation



Year of publication 2002
Type Article in Periodical
Magazine / Source Acta veterinaria (Brno)
Description With increasing importance of apoptosis, many assays have been developed to detect apoptosis. in cell populations and individual cells. Apoptotic cells are often proved using TUNEL assay based on detection of DNA strand breaks that occur during apoptosis. The method enables quantification of apoptotic cells by flow cytometry, fluorescence as well as by light microscopy when modifications using marker enzymes are applied. Disadvantages of this method are false positive and negative findings. In this study, the influence of different methods of fixation on TUNEL POD results was investigated, Three commonly employed fixative procedures using formaldehyde, methacarn and Bouin's fluid were applied. Two bovine tissue types of different cell density (thymus and ovary) were used to test this effect. Different pretreatments were tested for an influence upon false negative findings. Results were evaluated under light microscopy and photographed. We observed that, in the absence of a universal molecular marker of programmed cell death, it is essential for reliable apoptosis detection to optimize the chosen method for a given tissue and fixation. It is also strongly recommended to employ at least two methods, measuring different features of the apoptotic process to obtain reproducible results in detection of apoptosis.

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