A new fluorochromasia method using a fluorescence microplate reader for assay of cytotoxic activity of carp leucocytes.
A new fluorochromasia method using a fluorescence microplate reader has been established to assay spontaneous cytotoxic activity of carp leucocytes. This method is characterized by using propidium iodide (PI) for staining dead target cells and a fluorescence microplate reader for measurement of the fluorescence of PI. K562 as target cells were prepared in 96-well flat-bottom microplates, and carp leucocytes were added as effector cells. After 2.5 h incubation, PI was added to each well. After an additional 1.5 h incubation, fluorescence of each well was measured. Correlation between this method and 51Cr-release assay was obtained. The results demonstrated that this new fluorochromasia method can be used to assay cytotoxic activity of carp leucocytes.
Department of Aquatic Biosciences, Tokyo University of Fisheries, Japan.
N
N
Sano
O
O
Kurata
N
N
Okamoto
Y
Y
Ikeda
D000818:Animals; D002347:Carps / Q000276:immunology*; D003601:Cytotoxicity Tests, Immunologic / Q000379:methods / Q000662:veterinary*; D003602:Cytotoxicity, Immunologic / Q000276:immunology*; D005260:Female; D007962:Leukocytes / Q000276:immunology*; D008297:Male; D011419:Propidium; D013050:Spectrometry, Fluorescence / Q000662:veterinary
D011419:Propidium