Dual staining with Hoescht 33342 and PI

MORGANTL@aol.com
Tue, 3 Oct 1995 22:53:19 -0400

Has anyone had experience with the dual staining method described by Belloc
et al (Cytometry 17: 59-65; 1994)? We are using a B-D FACStar Plus with an
argon laser tuned to 310 nm. We are using the filters suggested by Belloc:
485/22 nm band pass and 600 nm long pass. As postive test cells we are are
using a Burkitt's lymphoma cell line (Ramos) treated with H7.

Our problem comes when we add both dyes together. We are able to detect blue
fluoroesence from the Hoescht 33342 alone. However, when we add PI, the
Hoescht channel PMT voltage has to be readjusted above 700 volts to achieve
even a minimal ratio signal.

Thanks in advance for any help and advice.

Thomas L. Morgan, Ph.D.
Regional Research Lab
Kaiser Permanente Medical Center
1515 N. Vermont Ave
Los Angeles, CA 90027
213-667-3560 213-667-3560 (FAX)