title: Rapid quantification of cell viability and apoptosis in B-cell lymphoma cultures using cyanine SYTO probes.
abstract: The gross majority of classical apoptotic hallmarks can be rapidly examined by multiparameter flow cytometry. As a result, cytometry became a technology of choice in diverse studies of cellular demise. In this context, a novel class of substituted unsymmetrical cyanine SYTO probes has recently become commercially available. Derived from thiazole orange, SYTO display low intrinsic fluorescence, with strong enhancement upon binding to DNA and/or RNA. Broad selection of excitation/emission spectra has recently driven implementation of SYTO dyes in polychromatic protocols with the detection of apoptosis being one of the most prominent applications In this chapter, we outline a handful of commonly used protocols for the assessment of apoptotic events using selected SYTO probes (SYTO 16, 62, 80) in conjunction with common plasma membrane permeability markers (PI, YO-PRO 1, 7-AAD).
authors.affiliation: The BioMEMS Research Group, Department of Chemistry, University of Auckland, Auckland 1142, New Zealand. d.wlodkowic@auckland.ac.nz
authors.forename: Donald
authors.initials: D
authors.lastname: Wlodkowic
authors.forename: Joanna
authors.initials: J
authors.lastname: Skommer
authors.forename: Zbigniew
authors.initials: Z
authors.lastname: Darzynkiewicz
mesh_terms: D017209:Apoptosis; D001583:Benzoxazoles / Q000378:metabolism; D002470:Cell Survival; D003584:Cytological Techniques / Q000379:methods*; D003609:Dactinomycin / Q000031:analogs & derivatives / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism*; D006801:Humans; D016393:Lymphoma, B-Cell / Q000473:pathology*; D015335:Molecular Probes / Q000378:metabolism*; D011419:Propidium / Q000378:metabolism; D011806:Quinolinium Compounds / Q000378:metabolism; D014407:Tumor Cells, Cultured
chemical_list: D001583:Benzoxazoles; D005456:Fluorescent Dyes; D015335:Molecular Probes; D011806:Quinolinium Compounds; C089813:YO-PRO 1; D003609:Dactinomycin; D011419:Propidium; C025942:7-aminoactinomycin D