Flow cytometric detection of saxitoxins using fluorescent voltage-sensitive dyes.
By virtue of their ability to block depolarization of nerve cells, the saxitoxins exert the toxic effects associated with paralytic shellfish poisoning and allow for their detection through various methodologies. When veratridine-induced depolarization is followed using voltage-sensitive fluorescent dyes, the presence of these toxic blocking agents can be observed as a decrease in fluorescence of dye-treated nerve cells. Detection using flow cytometry provides for selection of the most responsive population of cultured mouse neuroblastoma (Neuro 2a) cells thereby enhancing assay sensitivity and this approach can be accomplished in real time. The method is demonstrated in preliminary studies using saxitoxin and crude shellfish extracts.
Manger
Ronald
R
Fred Hutchinson Cancer Research Center, Biologics Production Facility, Seattle, WA 98109, USA. rmanger@fhcrc.org
Woodle
Doug
D
Berger
Andrew
A
Hungerford
James
J
D000818:Animals; D045744:Cell Line, Tumor; D005434:Flow Cytometry / Q000379:methods*; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry*; D049880:Mytilus edulis / Q000737:chemistry; D012530:Saxitoxin / Q000032:analysis / Q000737:chemistry*; D014701:Veratridine / Q000032:analysis / Q000737:chemistry
D005456:Fluorescent Dyes; D012530:Saxitoxin; D014701:Veratridine