Staining Methods for Programmed Cell Death
Formation and maintenance of the body plan is determined by several intrinsic factors including cell division, differentiation, and death. While structural and molecular markers for the detection of division and differentiation have long been used, detection of dying or dead cells is often more difficult to accomplish. This is mainly due to the fact that cell death can be genetically programmed (programmed cell death, PCD) or unprogrammed, and this distinction is often difficult to capture. This protocol presents a simple staining technique using trypan blue that can be utilized as a preliminary study to determine the presence (or absence) of dead cells within the tissue analyzed, but without discriminating between the type of death (programmed or unprogrammed). Once the presence of dead cells is verified, the more expensive and time-consuming terminal deoxynucleotidyl transferase (TdT) dUTP nick end labeling (TUNEL) analysis can be employed to localize cells undergoing PCD. Although straightforward in its steps, the presented protocol might require some optimizations depending on the type of the tissue used and/or its developmental stage.
KeywordsNecrosis Programmed cell death Terminal deoxynucleotidyl transferase (TdT) dUTP nick end labeling (TUNEL) Trypan blue
- 14.Rate DN, Cuenca JV, Bowman GR, Guttman DS, Greenberg JT (1999) The gain-of-function Arabidopsis acd6 mutant reveals novel regulation and function of the salicylic acid signaling pathway in controlling cell death, defenses, and cell growth. Plant Cell 11:1695–1708PubMedCentralCrossRefPubMedGoogle Scholar