3-Carboxy-6-chloro-7-hydroxycoumarin: A highly fluorescent, water-soluble violet-excitable dye for cell analysis

被引：28

作者：

Abrams, Barny ^{[1
]}

Diwu, Zhenjun ^{[2
]}

Guryev, Oleg ^{[1
]}

Aleshkov, Sergei ^{[3
]}

Hingorani, Ravi ^{[3
]}

Edinger, Mark ^{[1
]}

Lee, Rita ^{[1
]}

Link, Joe ^{[1
]}

Dubrovsky, Tim ^{[1
]}

机构：

[1] BD Biosci, Cell Anal, San Jose, CA 95131 USA

[2] ABDBioquest, Sunnyvale, CA 94085 USA

[3] BD Biosci, Cell Anal, San Diego, CA 92121 USA

来源：

ANALYTICAL BIOCHEMISTRY | 2009年 / 386卷 / 02期

关键词：

Cell analysis; Immunophenotyping; Violet-excitable dye; Multicolor flow cytometry; Lymphocytes; Subsetting; EXCITED FLUOROCHROMES; FLOW-CYTOMETRY; LASER-DIODES; SERIES; BRIGHT;

D O I：

10.1016/j.ab.2008.12.018

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

In our search for new violet-excitable dyes with improved photophysical and photochemical properties, we examined several halogen-substituted hydroxycoumarins and found that chlorinated derivatives are at least as bright as their fluorinated analogs. A monochlorinated hydroxycoumarin was found to have a high quantum yield (similar to 0.98), and human leucocyte-specific monoclonal antibodies (CD3, CD4. and CD45) conjugated with this dye exhibited reliable performance in flow cytometry assays. Additional studies were performed, with BD Horizon V450-antibody conjugates being included in eight-color cocktails aimed at subsetting lymphocytes and myeloid cells. Such cocktails can frequently be unstable due to the tendency of one or more components to lose structural integrity, photobleach, or develop unwanted affinities for another component, However, the cocktails employed in this study enabled several different applications to be run and established that multicolor reagent mixtures containing V450-antibody conjugates are functional and stable. (C) 2009 Elsevier Inc. All rights reserved.

引用

页码：262 / 269

页数：8

共 15 条

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