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荧光法测定组织培养板中培养细胞的DNA含量。

Fluorometric determination of the DNA content of cells cultured in tissue culture plates.

作者信息

Goppelt-Struebe M, Golombek M

机构信息

Institute of Molecular Pharmacology, Medical School Hannover, Germany.

出版信息

J Immunol Methods. 1992 Jul 6;151(1-2):245-8. doi: 10.1016/0022-1759(92)90123-b.

Abstract

Determining the DNA content of cultured cells with the fluorochrome diaminobenzoic acid (DABA) has been shown to be a sensitive and selective method. A procedure is described which applies this method to cells cultured in 96-well and 24-well plates. After removal of the medium, the cells are disrupted by sonication, dried and reacted with DABA. For the final read-out the DABA-DNA solution is transferred to standard cuvettes and the fluorescence intensity determined with a conventional fluorometer. The method is highly reproducible and suitable for the cell concentrations usually employed in microculture systems. It is an easy non-radioactive method for determining the numbers of both adherent and non-adherent cells, especially when comparing different stages of activation or differentiation.

摘要

用荧光染料二氨基苯甲酸(DABA)测定培养细胞的DNA含量已被证明是一种灵敏且具有选择性的方法。本文描述了一种将该方法应用于在96孔板和24孔板中培养的细胞的程序。去除培养基后,通过超声处理使细胞破碎,干燥并与DABA反应。为了进行最终读数,将DABA-DNA溶液转移至标准比色皿中,并用传统荧光计测定荧光强度。该方法具有高度可重复性,适用于微培养系统中通常使用的细胞浓度。它是一种简便的非放射性方法,可用于测定贴壁细胞和非贴壁细胞的数量,尤其是在比较激活或分化的不同阶段时。

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