ISSN:
1573-0603
Keywords:
CHSE-214
;
H33258
;
Cell proliferation
;
Calcium requirement
;
CytoFluor 2350
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Summary The use of a DNA-binding dye, bisbenzimidazole (H33258), and a microplate fluorometer, CytoFluor 2350, was optimized to measure cell number in Chinook salmon embryo cell cultures (CHSE-214). The uniformity of cell homogenates, which were prepared prior to staining, was evaluated by area scan, which consists of multiple measurements over the total well area. Disruption in 0.01% SDS produced a relatively uniform homogenate and a low background. Homogenates were stained with H33258 at 1 to 10 µg/ml. Linear relationships between cell numbers and fluorescence units were established in 96 well plates, which were read only by standard scan, and in 6, 12, 24 and 48 well plates, which were read by area and standard scan. Area scan provided better relationships. These methods were used to show that in L-15 medium CHSE-214 were able to attach, retain viability, and proliferate with very little exogenous calcium.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF01404822
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