Cy7PE and Cy7APC: Bright new probes for immunofluorescence
Article first published online: 6 DEC 1998
Copyright © 1996 Wiley-Liss, Inc.
Volume 24, Issue 3, pages 191–197, 1 July 1996
How to Cite
Roederer, M., Kantor, A. B., Parks, D. R. and Herzenberg, L. A. (1996), Cy7PE and Cy7APC: Bright new probes for immunofluorescence. Cytometry, 24: 191–197. doi: 10.1002/(SICI)1097-0320(19960701)24:3<191::AID-CYTO1>3.0.CO;2-L
- Issue published online: 6 DEC 1998
- Article first published online: 6 DEC 1998
- Manuscript Accepted: 29 MAR 1996
- Manuscript Received: 23 FEB 1996
- NIH. Grant Number: CA42509
- tandem dyes;
- resonance energy transfer;
- multiparameter measurements
We demonstrate the utility of indotricarbocyanine (Cy7) conjugates of the phycobiliproteins phycoerythrin (PE) and allophycocyanin (APC) in flow cytometry. This is the first demonstration of the use of an APC tandem dye for fluorescence measurements. These resonance energy transfer tandem dyes can be excited by the phycobiliprotein-specific excitation wavelengths and fluoresce at wavelengths above 780 nm. The tandem dyes, when conjugated to antibodies, are suitable for flow cytometry and other immunofluorescence applications. These conjugates are easily detectable above the very low autofluorescence in this part of the spectrum. Indeed, the Cy7-conjugated PE tandem (Cy7PE) has a “brightness” (fluorescence signal over cellular autofluorescence) comparable to that of fluorescein, and the Cy7APC tandem has a “brightness” comparable to that of APC. These tandems are also easily distinguished from other commonly used fluorophores, making them suitable for high-order multiparametric analysis. We show an example of six-color immunofluorescence analysis by flow cytometry, simultaneously measuring fluorescences from fluorescein, PE, Cy5PE, Texas red, APC, and Cy7APC. © 1996 Wiley-Liss, Inc.