Apoptosis Tissue sections

Gold, R, Schmied, M., Rothe, G, Zischler, H., Breitschopf, H, Wekerle, H, and Lassmann, H, (1993) Detection of DNA fragmentation in apoptosis- application of in situ nick translation to cell culture systems and tissue sections J Histochem Cytochem 41, 1023-1030... 

Immunohistochemistry, on the other hand, enables identification of activated caspases or their cleaved products in fixed archival tissue sections. This technique allows identification of cell(s) undergoing caspase activation, as well as analysis of the distribution of cell(s) in the tissue. Specific antibodies to various caspases are now commercially available, the most frequently studied being caspase-3. Studies in various human tissues and cells have shown that immunohistochemical detection of activated caspase-3 is a useful tool for identifying apoptotic cells in archival material, even before all of the morphological features of apoptosis occur [84-86]. Several target proteins cleaved by caspases can also be detected by immunohistochemistry for example PARP [87], actin [88, 89], and lamin B [90]. 

Many assays of apoptosis by LSC are performed on fixed cells. For these assays the cells are attached to microscope slides by standard methods that include smear films, tissue sections, touch preparations from freshly transected tissues, or cytocentrifuging cell suspensions. Cytocentrifugation (see below) is often preferred over touch or smear preparations because it flattens cells on the slides so that their geometry is favorable, and therefore more morphological details can be revealed. 

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