Flow cytometry instruments with solid state violet laser diodes are becoming more common due to their small size, low power requirements, cost, and reliability. The use of violet-excited fluorochromes in multiparametric flow cytometry has been essential in the expansion of multicolor flow cytometry, enabling greater numbers of markers to be detected in one sample. We have developed a novel violet-excited dye, Pacific Green™ dye, which has an excitation/emission maximum of 411 and 500 nm, respectively. This development addresses the need to transfer well-resolved markers off the Blue 488 nm and Red 635 nm excitation lines and onto the Violet 405 nm excitation line, thus enabling the detection of other markers with the 488 nm and 635 nm lasers. Pacific Green™ dye can be used for three color immunophenotyping using violet laser excitation with Pacific Blue™ and Pacific Orange ™ dyes with minimal compensation without 488 nm excitation. Data is shown for human lymphocytes and mouse splenocytes stained with Pacific Green™ direct conjugates. Six color immunophenotyping data is shown using direct conjugates of Pacific Green™, Alexa Fluor® 488, phycoerythrin, PE-Cy®7, Pacific Blue™ and Pacific Orange™ conjugates. Pacific Green™ dye can be used with the far red emitting QDot®605, 655, and 705 conjugates for higher multiplexing with violet laser excitation. Pacific Green™ dye can be paired with fixable blue or yellow-emitting viability stain with violet excitation to exclude dead cells.
- Copyright © 2013 by The American Association of Immunologists, Inc.