Journal
CYTOMETRY PART A
Volume 73A, Issue 6, Pages 570-577Publisher
WILEY-LISS
DOI: 10.1002/cyto.a.20563
Keywords
yellow laser; orange laser; diode pumped solid state; red fluorescent protein; Texas Red; allophycocyanin; HcRed; Katushka
Categories
Funding
- Intramural NIH HHS [ZIC BC011138] Funding Source: Medline
- NIGMS NIH HHS [GM073913, R01 GM070358, GM070358, R01 GM073913] Funding Source: Medline
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Diode and DPSS lasers emitting a variety of wavelengths are now commonly incorporated into flow cytometers, greatly increasing our capacity to excite a wide variety of fluorochromes. Until recently, however, virtually no practical technology existed for generating yellow or orange laser light for flow cytometry that was compatible with smaller instrumentation. In this study, we evaluate several new solid state laser systems that emit from the 570 to 600 nm as excitation sources for flow cytometry. DPSS 580, 589, and 592 nm sources were integrated into a cuvette-based flow cytometer (BD LSR II) and a stream-in-air cell sorter (FACSVantage DiVa), and used to excite a variety of yellow, orange, and red excited fluorochromes, including Texas Red, APC, and its tandem conjugates, and the genetically encoded red fluorescent protein HcRed and the more recently developed Katushka. All laser sources were successfully incorporated into the indicated flow cytometry platforms. The yellow and orange sources (particularly 592 nm) were ideal for exciting Texas Red, and provided excitation of APC and its tandems that was comparable to a traditional red laser source, albeit at higher power levels than red sources. Yellow and orange laser light was optimal for exciting HcRed and Katushka. Practical yellow and orange laser sources are now available for flow cytometry. This technology fills an important gap in the laser wavelengths available for flow, now almost any fluorochrome requiring visible light excitation can be accommodated. Published 2008 Wiley-Liss, Inc.
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