SIMPLIFIED AND OPTIMIZED KINETOCHORE DETECTION - CYTOGENETIC MARKER FOR LATE-G(2) CELLS

被引：7

作者：

ABEND, M ^{[1
]}

BLAKELY, WF ^{[1
]}

VANBEUNINGEN, D ^{[1
]}

机构：

[1] ARMED FORCES RADIOBIOL RES INST,DEPT RADIAT BIOPHYS,BETHESDA,MD 20889

来源：

MUTATION RESEARCH-ENVIRONMENTAL MUTAGENESIS AND RELATED SUBJECTS | 1995年 / 334卷 / 01期

关键词：

KINETOCHORES; LATE-G; PHASE; FLUOROCHROME CONJUGATION; CY3; BODIPY; FITC;

D O I：

10.1016/0165-1161(95)90029-2

中图分类号：

X [环境科学、安全科学];

学科分类号：

08 ; 0830 ;

摘要：

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-X100, 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 kinetochores, the latter of which are characteristic of late-G(2) phase and prophase cells.

引用

页码：39 / 47

页数：9

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