Get 20M+ Full-Text Papers For Less Than $1.50/day. Start a 14-Day Trial for You or Your Team.

Learn More →

ALLO‐7: A new fluorescent tandem dye for use in flow cytometry

ALLO‐7: A new fluorescent tandem dye for use in flow cytometry This report describes the development of a novel tandem dye by combining allophycocyanine (APC) and cyanine dye indotricarbocyanine (CY7) to create ALLO‐7 for use in flow cytometry. The APC donor fluorophore was excited at 647 nm and, through resonance energy transfer to the CY7 acceptor, produced fluorescence at >780 nm. To test the applicability of this tandem in single and multicolor immunofluorescence, a streptavidin conjugate of the tandem (SA‐ALLO‐7) was used for the detection of cell surface antigens on human peripheral blood leukocytes (PBL) by indirect immunofluorescence. Human PBL were stained with CD4/GaM‐APC, CD3‐fluorescein isothiocyanate (FITC), CD14‐phycoerythrin (PE), CD19‐energy‐coupled dye (phycoerythrin‐Texas Red) (ECD), and CD8‐biotin with SA‐ALLO‐7 and analyzed for fluorescence on a FACS Vantage using dual‐laser excitation (488 and 647 nm). The results indicated that the percentage of cells positive for each of the surface antigens was comparable for single‐color controls and multicolor samples. The ALLO‐7 fluorescence, which was collected with a 730‐shortpass dichroic mirror and a 790/50‐bandpass filter, was clearly resolved from the APC fluorescence and that from FITC, PE, and ECD. The SA‐ALLO‐7 exhibited minimal nonspecific binding to PBL monocytes. However, the specific binding of the tandem to high‐density antigens was clearly identified by positive fluorescence. This unique tandem reagent, ALLO‐7, provided the capability for dual‐color immunofluorescence with a 647‐nm laser line (or a helium neon laser at 633 nm) and provides the potential to perform three‐color analysis with a dye‐head laser (Texas Red, APC, ALLO‐7). © 1996 Wiley‐Liss, Inc. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Cytometry Wiley

ALLO‐7: A new fluorescent tandem dye for use in flow cytometry

Beavis, Andrew J.; Pennline, Kenneth J.
Cytometry , Volume 24 (4) – Aug 1, 1996
Download PDF
5 pages

Loading next page...
 
/lp/wiley/allo-7-a-new-fluorescent-tandem-dye-for-use-in-flow-cytometry-8qhe0Qxc21

References (0)

References for this paper are not available at this time. We will be adding them shortly, thank you for your patience.

Publisher
Wiley
Copyright
Copyright © 1996 Wiley‐Liss, Inc., A Wiley Company
ISSN
0196-4763
eISSN
1097-0320
DOI
10.1002/(SICI)1097-0320(19960801)24:4<390::AID-CYTO11>3.3.CO;2-L
Publisher site
See Article on Publisher Site

Abstract

This report describes the development of a novel tandem dye by combining allophycocyanine (APC) and cyanine dye indotricarbocyanine (CY7) to create ALLO‐7 for use in flow cytometry. The APC donor fluorophore was excited at 647 nm and, through resonance energy transfer to the CY7 acceptor, produced fluorescence at >780 nm. To test the applicability of this tandem in single and multicolor immunofluorescence, a streptavidin conjugate of the tandem (SA‐ALLO‐7) was used for the detection of cell surface antigens on human peripheral blood leukocytes (PBL) by indirect immunofluorescence. Human PBL were stained with CD4/GaM‐APC, CD3‐fluorescein isothiocyanate (FITC), CD14‐phycoerythrin (PE), CD19‐energy‐coupled dye (phycoerythrin‐Texas Red) (ECD), and CD8‐biotin with SA‐ALLO‐7 and analyzed for fluorescence on a FACS Vantage using dual‐laser excitation (488 and 647 nm). The results indicated that the percentage of cells positive for each of the surface antigens was comparable for single‐color controls and multicolor samples. The ALLO‐7 fluorescence, which was collected with a 730‐shortpass dichroic mirror and a 790/50‐bandpass filter, was clearly resolved from the APC fluorescence and that from FITC, PE, and ECD. The SA‐ALLO‐7 exhibited minimal nonspecific binding to PBL monocytes. However, the specific binding of the tandem to high‐density antigens was clearly identified by positive fluorescence. This unique tandem reagent, ALLO‐7, provided the capability for dual‐color immunofluorescence with a 647‐nm laser line (or a helium neon laser at 633 nm) and provides the potential to perform three‐color analysis with a dye‐head laser (Texas Red, APC, ALLO‐7). © 1996 Wiley‐Liss, Inc.

Journal

Cytometry – Wiley

Published: Aug 1, 1996

Keywords: Tandem dye; CY7; APC; multicolor analysis; flow cytometry; far red emission

There are no references for this article.