Accession Number : ADA293794
Title : AFRRI Reports, First Quarter 1995, January - March 1995.
Corporate Author : ARMED FORCES RADIOBIOLOGY RESEARCH INST BETHESDA MD
PDF Url : ADA293794
Report Date : 1995
Pagination or Media Count : 100
Abstract : This volume contains AFRRI Scientific Reports SR95-1 through SR95-10 for January-March 1995. Cytogenetic detection of kinetochore proteins using the CREST antibody coupled with secondary antibodies labeled with different fluorescent probes has been optimized for several in vitro mammalian cell lines. This study investigated selected parameters including the influence of common fixatives (methanol, ethanol, methanol : acetic acid (3: 1)), detergents (Triton-XlOO, Tween), fluorescent probes (CY3, BODIPY, FITC), washing protocols, and an antifading agent (paraphenylenediamine) on the detection of kinetochore proteins in control and X-ray (240 kVp)-irradiated cells. Utilizing an optimized fixation and staining protocol, a brilliant visualization of kinetochores in interphase cells was obtained in control as well as X-ray-irradiated interhase cells. Application of this improved kinetochore staining methodology readily permits discriminating cells containing either single or paired kineto- chores, the latter of which are characteristic of late-G2 phase and prophase cells.
Descriptors : *FLUORESCENCE, *METHANOLS, *ANTIBODIES, PROBES, DETECTION, CELLS, SECONDARY, PROTEINS, PHASE STUDIES, X RAYS, DETERGENTS, IRRADIATION, DISCRIMINATION, ACETIC ACID.
Subject Categories : Medicine and Medical Research
Distribution Statement : APPROVED FOR PUBLIC RELEASE