Detection of Apoptosis

D5. Donner, K. J., Becker, K. M., Hissong, B. D., and Ahmed, S. A., Comparison of multiple assays for kinetic detection of apoptosis in thymocytes exposed to dexamethasone or diethylstilbesterol. 

V2. Van den Eijnde, S. M., Luijsterburg, A. J., Boshart, L. De Zeeuw, C. I., van Dierendonck, J. H., Reutelingsperger, C. R, and Vermeij-Keers, C., In situ detection of apoptosis during embryogenesis with annexin V From whole mount to ultrastructure. Cytometry 29, 313-320 (1997). 

Swat, W., lgnatowicz, L., and Kisielow, P. (1991) Detection of apoptosis of immature CD4+8+ thymocytes by flow cytometry. J. Immunol. Methods 137,79-87. 

Mochizuki H, Goto K, Mori H, Mizuno Y. 1996. Histochemical detection of apoptosis in Parkinson s disease. J Neurol Sci 137 120-123. 

Despite its wide use, the specificity of the TUNEL method for detection of apoptosis has been seriously questioned. There is mounting evidence that TUNEL is unreliable for the identification of apoptosis if used alone. DNA strand breaks are not a unique feature of apoptosis, since they have been found in various tissues, also in necrosis, during repair of reversibly damaged DNA, active gene transcription, and postmortem autolysis [85, 96-98],... 

Apoptosis has been demonstrated to occur also in human MI. Studies of heart tissue obtained at autopsies from patients, who died of MI, suggest that apoptotic cardiomyocytes were most prominent in the border zones of MI (Fig. 1A and B) [111-123], whereas few were present in the remote myocardium [113, 114, 118-121]. The most commonly used method for detection of apoptosis in these studies was TUNEL. Using this method, a highly variable apoptotic rate was found, ranging from 0.8% [114] to 43% [118] in the border zone of MI, and from 0.05% [114] to 38% [118] in the remote myocardium. Using immunohistochemistry for activated (cleaved) caspase-3, a much lower apoptotic rate was reported in human MI [123]. 

Rodriguez-Calvo MS, Tourret MN, Concheiro L, Munoz JI, Suarez-Penaranda JM. Detection of apoptosis in ischemic heart. Usefulness in the diagnosis of early myocardial injury. Am J Forensic Med Pathol 2001 22 278-284. 

G6. Gorczyca, W., Tuziak, T., Kram, A., Melamed, M. R., and Darzynkiewicz, Z., Detection of apoptosis-associated DNA strand breaks in fine-needle aspiration biopsies by in situ end labelling of fragmented DNA. Cytometry 15, 169-175 (1994). 

Examples of Detection of Apoptosis with a Confocal Microsocope... 

Telford, W. G., King, L. E., and Fraker, P. J. (1992) Comparative evaluation of several DNA binding dyes in the detection of apoptosis-associated chromatin degradation by flow cytometry. Cytometry 13(2), 137-143. 

Hotz, M. A., Gong, J., Traganos, F., and Darzynkiewicz, Z. (1994) Flow cytometric detection of apoptosis comparison of the assays of in situ DNA degradation and chromatin changes. Cytometry 15(3), 237-244. 

Poot, M. and Pierce, R. C. (1999) Detection of apoptosis and changes in mitochondrial membrane potential with chloromethyl-X-rosamine [letter]. 

Li, X. and Darzynkiewicz, Z. (1995) Labelling DNA strand breaks with BrdUTP. Detection of apoptosis and cell proliferation. Cell Prolif. 28, 571-579. 

USE OF OTHER APOPTOSIS-REGULATED PROTEINS AND EVENTS IN DETECTION OF APOPTOSIS... 

Mammalian. - The use of NMR imaging and spectroscopy for the detection of apoptosis has been reviewed. ... 

K., and Ishii, T. (2001) Detection of Apoptosis in Keloids and a Comparative Study on Apoptosis Between Keloids, Hypertrophic Scars, Normal Healed Flat Scars and Dermatofibroma, Wound Repair Regen. 9,501-506. 

Ohtsuki K, Akashi K, Aoka Y, Blankenberg FG, Kopiwoda S, Tait JF, Strauss HW. Technetium-99m HYNIC-annexin V a potential radiopharmaceutical for the in vivo detection of apoptosis. Eur J Nucl Med 1999 26 1251 1258. 

Frey, T. Nucleic acid dyes for detection of apoptosis in live cells. Gytometry 1995, 21, 265-274. 

Matsuo, A. Watanabe, A. Takahashi, T. Futamura, M. Mori, S. Sugiyama, Y. Takahashi, Y Saji, S. A simple method for classification of cell death by use of thin layer collagen gel for the detection of apoptosis and/or necrosis after cancer chemotherapy. Jpn. J. Cancer Res. 2001, 92, 813-819. 

Bossy-Wetzel, E. Green, D. R. Detection of apoptosis by annexin V labeling. Methods Enzymol. 2000, 322, 15-18. 

Reynolds, J. E. Li, J. Eastman, A. Detection of apoptosis by flow cytometry of cells simultaneously stained for intracellular pH (carboxy SNARE-1) and membrane permeability (Hoechst 33342). Cytometry 1996, 25, 349-357. 

Wlodkowic, D. Skommer, J. Hillier, C. Darzynkiewicz, Z. Multiparameter detection of apoptosis using red-excitable SYTO probes. 

Willingham MC (1999) Cytochemical methods for the detection of apoptosis. J Histochem Cytochem 47 1101-1109... 

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