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不同批次金胺O(OO)的光谱和光度特性

[Spectral and photometric characteristics of different batches of auramine O (OO)].

作者信息

Kudriavtsev B N, Rautian L P, Kudriavtseva M V, Rozanov Iu M

出版信息

Tsitologiia. 1975 Sep;17(9):1103-8.

PMID:52224
Abstract

The spectra of absorption and fluorescence of Auramine O (OO) specimens in water solution and in cells, stained with Schiff-type auramine-SO2 reagents for DNA, were investigated. Using different specimens of the stain the accuracy and sensitivity of the cytofluorometric method were estimated by the next parameters: the value of the useful signal from the nucleus (no), the value of the production of no by the time constant of fading (tau), the value of proportion of the signal to the background (no/nphi) and, in addition, the value of the variation coefficient under the measurement of the rat liver tetraploid cells. The purity of the dye was shown to interfere largely with accuracy and sensitivity of the above method. Auramine 00 of the Reanal production appeared to have the best photometricl characteristics. For this, the maxima of the absorption spectrum in the 320--700 nm region in the solution and in the cell were 371 and 433 nm, and 375 and 436 nm, resp. The maximum of the fluorescence spectrum was found 521 nm and 526 nm, resp. for the solution (0.02%) and for the Auramine-SO2 treated cells. Moreover, an isomer of Auramine O (00) -- Auramine G can be used for detection and photometrical measurement of DNA and glycogen after the Feulgen and PAS reactions, resp.

摘要

研究了水溶液和细胞中碱性槐黄O(OO)标本的吸收光谱和荧光光谱,这些标本用用于DNA染色的席夫型碱性槐黄-SO₂试剂进行了染色。使用不同的染色标本,通过以下参数评估细胞荧光测定法的准确性和灵敏度:来自细胞核的有用信号值(no)、no随褪色时间常数(tau)的产生值、信号与背景的比例值(no/nphi),此外,还有大鼠肝脏四倍体细胞测量时的变异系数值。结果表明,染料的纯度在很大程度上会干扰上述方法的准确性和灵敏度。Reanal生产的碱性槐黄OO似乎具有最佳的光度特性。为此,溶液和细胞中320 - 700 nm区域吸收光谱的最大值分别为371和433 nm,以及375和436 nm。荧光光谱的最大值分别为521 nm和526 nm,分别对应于溶液(0.02%)和经碱性槐黄-SO₂处理的细胞。此外,碱性槐黄O(OO)的一种异构体——碱性槐黄G可分别用于费尔根反应和PAS反应后DNA和糖原的检测和光度测量。

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