期刊
JOURNAL OF NEUROSCIENCE METHODS
卷 100, 期 1-2, 页码 157-163出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/S0165-0270(00)00248-X
关键词
neurotoxicity; trypan blue; lactate dehydrogenase; cell culture; cortical neurons; N-methyl-D-aspartate; anoxia; spectrophotometry
资金
- NINDS NIH HHS [NS36812, R01 NS036812] Funding Source: Medline
An automated method for the determination of neuronal cell death using trypan blue is described. Following various excitotoxic insults, murine mixed cortical cell cultures are stained with trypan blue (0.05%; 15 min), followed by SDS (1%) lysis. The absorbance of the dye is measured spectrophotometrically at 590 nm using a microtiter plate reader. When compared to the biochemical lactate dehydrogenase assay, no statistical difference in the calculated levels of excitotoxic neuronal cell death was noted between the assays in ally given paradigm. This method is fast and reliable. It eliminates the need for cell counting, thus allowing for high volume sample analysis with a minimum of sample error. Utility of this trypan blue absorbance spectrophotometric assay is likely to extend beyond the study of excitotoxic neuronal injury and should complement existing methods for measuring neuronal viability and cytotoxicity in cell culture. (C) 2000 Elsevier Science B.V. All rights reserved.
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