By Myrtle A. Davis
Dr. Myrtle A. Davis has assembled a panel of state of the art scientists to explain their top tools for detecting, illuminating, and quantifying apoptotic mechanisms in a manner that's beneficial for the layout of toxicology and pharmacology reports. those state of the art recommendations comprise move cytometric, fluorometric, and laser scanning tools for quantifying and characterizing apoptosis, in addition to protocols for using DNA microarray know-how, excessive throughput displays, and ELISA. Immunocytochemical equipment for measuring biochemical and molecular endpoints in tissue sections could be hugely worthy for these undertaking stories in complete animal types in place of cellphone tradition structures.
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Additional info for Apoptosis Methods in Pharmacology and Toxicology: Approaches to Measurement and Quantification (Methods in Pharmacology and Toxicology)
Cytometry 35(3), 181–195. 68. Fadok, V. , Voelker, D. , Campbell, P. , Cohen, J. , Bratton, D. , and Henson, P. M. (1992) Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages. J. Immunol. 148(7), 2207–2216. 69. van Heerde, W. , Doevendans, P. , Smits, J. , et al. CP (2000). Markers of apoptosis in cardiovascular tissues: focus on Annexin V. Cardiovasc. Res. 45(3), 549–559. 70. , Nieland, L. , Ramaekers, F. , and Reutelingsperger, C.
1999) Confocal microscopy of the mitochondrial permeability transition in necrotic and apoptotic cell death. Biochem. Soc. Symp. 66, 205–222. 23. Halestrap, A. P. (1999) The mitochondrial permeability transition: its mo lecular mechanism and role in reperfusion injury. Biochem. Soc. Symp. 66, 181–203. 24. , Susin, S. , and Kroemer, G. (1996) Inhibitors of permeability transition interfere with 32 25. 26. 27. 28. 29. 30. 31. 32. 33. 34. 35. 36. 37. Poot et al. the disruption of the mitochondrial transmembrane potential during apoptosis.
4. Set up and optimize the flow cytometer. UV-excited (360 nm) blue autofluorescence (collected with a 450 nm centered bandpass filter) is proportional with cellular NAD(P)H and to cellular reduced thiol content (in the samples stained with monobromobimane). To excite MitoTracker Green FM and CMXRosamine stained samples, the argon-ion laser should be tuned to the 488 nm line. To collect fluorescence from MitoTracker Green FM use a bandpass filter centered around 530 nm; for CMXRos use a longpass filter above 630 nm.
Apoptosis Methods in Pharmacology and Toxicology: Approaches to Measurement and Quantification (Methods in Pharmacology and Toxicology) by Myrtle A. Davis