Classification, Scoring, and Quantification of Cell Death in Tissue Sections

The analysis and description of the appearance of cell death in tissue sections can add valuable information to research studies. The scoring/grading and quantification of cell death can be used either as part of a larger scoring scheme or as the final end point of a study. The degree of precision needed is study dependent and will be determined by the question being addressed and the complexity of the model. The methods one uses to quantify cell death are often guided by the tissue of interest. For example, in the brain, it is sometimes necessary to examine death of specific neuronal populations, whereas in more homogeneous tissue such as a tumor xenograft, quantification can be done on a whole-slide basis. In addition to examination of hematoxylin and eosin (HE)-stained sections, immunohistochemistry can be employed to highlight areas of cell death or to identify specific types of cell death, for example, when differentiating apoptosis from necrosis. Automated quantification can be useful in generating statistically comparable data from HE-stained or immunolabeled samples. The rapidly expanding classification of cell death requires the use of multiple techniques to identify them in vivo. This article will provide examples of how different methods of examining and quantifying cell death are used in a variety of research areas, ranging from semiquantitative evaluation in HE-stained intestine to automated quantification of immunohistochemistry-immunolabeled brain and tumor xenografts. The recently described process of necroptosis will be discussed briefly, with the description and example of the methods used to differentiate this from apoptosis.