Open AccessVisible diode lasers can be used for flow cytometric immunofluorescence and DNA analysis
Author(s) -
Doornbos Richard M. P.,
de Grooth Bart G.,
Kraan Yvonne M.,
van der Poel Carel J.,
Greve Jan
Publication year - 1994
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/cyto.990150312
Subject(s) - immunofluorescence , flow cytometry , laser , diode , optoelectronics , optics , materials science , microbiology and biotechnology , biology , physics , genetics , antibody
This report describes a feasibility study concerning the use of a visible diode laser for two important fluorescence applications in a flow cytometer. With a 3 mW 635 nm. diode laser, we performed immunofluorescence measurements using the fluorophore allophycocyanin (APC). We have measured CD8 positive lymphocytes with a two‐step labeling procedure and the resulting histograms showed good separation between the negative cells and the dim and the bright fluorescent subpopulations. As a second fluorescence application, we chose DNA analysis with the recently developed DNA/ RNA stains TOTO‐3 and TO‐PRO‐3. In our setup TO‐PRO‐3 yielded the best results with a CV of 3.4%. Our results indicate that a few milliwatts of 635 nm light from a visible diode laser is sufficient to do single color immunofluorescence measurements with allophycocyanin and DNA analysis with TO‐PRO‐3. The major advantages of using a diode laser in a flow cytometer are the small size, the low price, the high efficiency, and the long lifetime. © 1994 Wiley‐Liss, Inc.