Scientific Publications - Work Done by Microbiology Reader Bioscreen C

Fiala J., Neumajer T. and Pichova A., Controlling cell proliferation and cell death (apoptosis) by flow cytometry, Web publication, 2003, Institute of Chemical Technology Prague, Department of Fermentation Chemistry and Bioengineering, Prague, Czech Republic; University of Salzburg, Department of Genetics, Salzburg, Austria

INTRODUCTION

Apoptosis is a carefully regulated process of cell death that occurs as a normal part of development. Apoptosis is distinguished from necrosis, or accidental cell death, by characteristic morphological and biochemical changes, including compaction and fragmentation of the nuclear chromatin, shrinkage of the cytoplasm and loss of membrane asymmetry. Annexin V conjugated to fluorescein (FITC annexin V) can identify apoptotic cells by binding to phosphatidyl serine (PS) and red-fluorescent propidium iodide (PI) bind nucleic acid of necrotic cells. These populations can be distinguished using a flow cytometer (Partec PAS III) with the 488 nm line of an argon-ion laser for excitation. We have optimised this assay using yeast cells - Saccharomyces cerevisiae (wild type (wt), RAS2val19, RAS2ser33, 2+a, MUT[155], R2-, MUT[F-]). Apoptosis is a carefully regulated process of cell death that occurs as a normal part of development. Apoptosis is distinguished from necrosis, or accidental cell death, by characteristic morphological and biochemical changes, including compaction and fragmentation of the nuclear chromatin, shrinkage of the cytoplasm and loss of membrane asymmetry. Annexin V conjugated to fluorescein (FITC annexin V) can identify apoptotic cells by binding to phosphatidyl serine (PS) and red-fluorescent propidium iodide (PI) bind nucleic acid of necrotic cells. These populations can be distinguished using a flow cytometer (Partec PAS III) with the 488 nm line of an argon-ion laser for excitation. We have optimised this assay using yeast cells - Saccharomyces cerevisiae (wild type (wt), RAS2val19, RAS2ser33, 2+a, MUT[155], R2-, MUT[F-]).

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Last modified: May 25, 2005