Two-color fluorescence in flow cytometry DNA sizing: Identification of single molecule fluorescent probes

被引：20

作者：

Agronskaia, A ^{[1
]}

Schins, JM ^{[1
]}

de Grooth, BG ^{[1
]}

Greve, J ^{[1
]}

机构：

[1] Univ Twente, Dept Appl Phys, Appl Opt Grp, NL-7500 AE Enschede, Netherlands

来源：

ANALYTICAL CHEMISTRY | 1999年 / 71卷 / 20期

关键词：

D O I：

10.1021/ac990422v

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

We have built a two laser flow cytometer for ultrasensitive fluorescence measurements. The instrument is used for two-parameter analysis of DNA fragments. Total DNA content of each fragment (proportional to its size) is measured using TOTO-1 excited at 514 nm. Specific targets on the DNA are labeled with Cy5 and excited at 647 nm. The sensitivity of the instrument allows the detection of targets at the single molecule level. The signal-to-noise ratio for a single streptavidin-conjugated Cy5 molecule (about four Cy5 molecules) is 7.5. The SNR for a single Cy5 molecule is 1.7.

引用

页码：4684 / 4689

页数：6

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