基于玻片的多色细胞术荧光染料的顺序光漂白
Sequential photobleaching of fluorochromes for polychromatic slide-based cytometry.
作者信息
Mittag Anja, Lenz Dominik, Bocsi József, Sack Ulrich, Gerstner Andreas O H, Tárnok Attila
机构信息
Department of Pediatric Cardiology, Heart Center Leipzig GmbH, University of Leipzig, Leipzig, Germany.
出版信息
Cytometry A. 2006 Mar;69(3):139-41. doi: 10.1002/cyto.a.20227.
BACKGROUND
Slide-based cytometry is a key technology for polychromatic cytomic investigations. Here we exploit the relocalization and merge feature of Laser Scanning Cytometry for distinguishing fluorochromes of comparable emission spectra but different photostabilities.
METHODS
Blood specimens were stained with the fluorochrome pairs: FITC/ALEXA488, PE/ALEXA532, or APC/ALEXA633. Bleaching was performed by repeated laser excitation.
RESULTS
Since ALEXA dyes are photostable as compared to the conventional fluorochromes FITC, PE, and APC, a differentiation within one fluorochrome pair is possible.
CONCLUSION
The sequential photobleaching method results in an increased information density on a single cell level and represents an important component to perform polychromatic cytometry.
背景
基于玻片的细胞计数法是多色细胞组学研究的关键技术。在此，我们利用激光扫描细胞计数法的重新定位和合并功能来区分发射光谱相近但光稳定性不同的荧光染料。
方法
血液标本用荧光染料对进行染色：异硫氰酸荧光素/藻红蛋白 Alexa488、藻红蛋白/藻红蛋白 Alexa532 或别藻青蛋白/藻红蛋白 Alexa633。通过重复激光激发进行漂白。
结果
由于与传统荧光染料异硫氰酸荧光素、藻红蛋白和别藻青蛋白相比，藻红蛋白染料具有光稳定性，因此在一个荧光染料对内进行区分是可能的。
结论
顺序光漂白法可提高单细胞水平的信息密度，是进行多色细胞计数的重要组成部分。
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