Apoptosis Methods in Pharmacology and Toxicology: Approaches by Myrtle A. Davis

By Myrtle A. Davis

Dr. Myrtle A. Davis has assembled a panel of state of the art scientists to explain their most sensible equipment for detecting, illuminating, and quantifying apoptotic mechanisms in a fashion that's important for the layout of toxicology and pharmacology experiences. those cutting-edge thoughts contain circulate cytometric, fluorometric, and laser scanning tools for quantifying and characterizing apoptosis, in addition to protocols for using DNA microarray know-how, excessive throughput monitors, and ELISA. Immunocytochemical equipment for measuring biochemical and molecular endpoints in tissue sections should be hugely beneficial for these undertaking reports in entire animal versions rather than telephone tradition platforms.

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Additional resources for Apoptosis Methods in Pharmacology and Toxicology: Approaches to Measurement and Quantification (Methods in Pharmacology and Toxicology)

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The data can be used to map the sequence of both functional and structural changes occurring during apoptosis to determine whether a particular death-associated event is a prerequisite for the latter steps. Applications for studies of apoptosis that descend from these unique features of LSC are presented further in this chapter. 3. ATTACHMENT OF CELLS TO MICROSCOPE SLIDES FOR ANALYSIS BY LSC 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.

Campbell, J. , Stephenson, A. , Franklin, C. , et al. (2000) Disruption of redox homeostasis in tumor necrosis factor-induced apoptosis in a murine hepatocyte cell line. Am. J. Pathol. 157(1), 221–236. Apoptotic Cell Death 33 38. Keij, J. , and Steinkamp, J. A. (2000) Staining of mitochondrial membranes with 10-nonyl acridine orange, MitoFluor Green, and MitoTracker Green is affected by mitochondrial membrane potential altering drugs. Cytometry 39(3), 203–210. 39. , Shen, S. , et al. (2000) Caspase-8-mediated intracellular acidification precedes mitochondrial dysfunction in somatostatin-induced apoptosis.

Cytometry 36(4): 359–360. 44. Ublacker, G. , Johnson, J. , Siegel, F. , and Mulcahy, R. T. (1991) Influence of glutathione S-transferases on cellular glutathione determination by flow cytometry using monochlorobimane. Cancer Res. 51(7), 1783–1788. 45. Thorell, B. (1983) Flow-cytometric monitoring of intracellular flavins simultaneously with NAD(P)H levels. Cytometry 4(1), 61–65. 46. Makrigiorgos, G. , Kassis, A. , Bump, E. , and Savvides, P. (1997) Novel fluorescein-based flow-cytometric method for detection of lipid peroxidation.

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