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

By Myrtle A. Davis

ISBN-10: 0896038904

ISBN-13: 9780896038905

ISBN-10: 1592592791

ISBN-13: 9781592592791

Dr. Myrtle A. Davis has assembled a panel of state of the art scientists to explain their most sensible tools for detecting, illuminating, and quantifying apoptotic mechanisms in a fashion that's priceless for the layout of toxicology and pharmacology stories. those state of the art ideas contain stream cytometric, fluorometric, and laser scanning tools for quantifying and characterizing apoptosis, in addition to protocols for using DNA microarray expertise, excessive throughput displays, and ELISA. Immunocytochemical equipment for measuring biochemical and molecular endpoints in tissue sections can be hugely beneficial for these undertaking stories in entire animal types in place of telephone tradition platforms.

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

Example text

Set up and optimize the flow cytometer as in Basic Protocol 1. UV-excited (360 nm) blue fluorescence (collected with a 450 nm centered bandpass filter) from the Hoechst 33342 dye is proportional with cellular DNA content. To excite MitoTracker Green FM and CMXRosamine stained samples, the argonion 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 (see Notes 5–8).

63. Endresen, P. , Prytz, P. , and Aarbakke, J. (1995) A new flow cytometric method for discrimination of apoptotic cells and detection of their cell cycle specificity through staining of F-actin and DNA. Cytometry 20(2), 162–171. 64. Leers, M. , et al. (1999) Immunocytochemical detection and mapping of a cytokeratin 18 neo- epitope exposed during early apoptosis. J. Pathol. 187(5), 567–572. 65. Morsi, H. , Leers, M. , Kabarity, H. , and Jager, W. (2000) Apoptosis, bcl-2 expression, and proliferation in benign and malignant endometrial epithelium: An approach using multiparameter flow cytometry.

Examples of Detection of Apoptosis with a Confocal Microsocope As with flow cytometry, multiparameter apoptosis assays may also be performed by confocal laser scanning microscopy (CLSM). Using the approach similar to that detailed above for flow cytometry, we have examined NADPH content, mitochondrial membrane potential (CMX Rosamine fluorescence), and mitochondrial mass (Mitotracker Green), by CLSM. Figure 3 shows an example of a typical multiparameter assay performed by confocal microscopy. One advantage to examining apoptotic cells by laser-scanning cytometry is that subcellular structure may be examined.

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Apoptosis Methods in Pharmacology and Toxicology: Approaches to Measurement and Quantification (Methods in Pharmacology and Toxicology) by Myrtle A. Davis

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