SELECTIVE LABELING OF EMBRYONIC NEURONS CULTURED ON ASTROCYTE MONOLAYERS WITH 5(6)-CARBOXYFLUORESCEIN DIACETATE (CFDA)

被引：39

作者：

PETROSKI, RE ^{[1
]}

GELLER, HM ^{[1
]}

机构：

[1] UNIV MED & DENT NEW JERSEY, ROBERT WOOD JOHNSON MED SCH, DEPT PHARMACOL, PISCATAWAY, NJ 08854 USA

来源：

JOURNAL OF NEUROSCIENCE METHODS | 1994年 / 52卷 / 01期

关键词：

NEURON-SPECIFIC MARKER; ASTROCYTE; IMMUNOCYTOCHEMISTRY; VITAL DYE;

D O I：

10.1016/0165-0270(94)90051-5

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A method for selectively labeling cultured neurons using the vital dye, 5(6)-carboxyfluorescein diacetate (CFDA), is described. This non-fluorescent membrane-permeant dye is cleaved by cytosolic esterases into the fluorescent anion, 5(6)-carboxyfluorescein (CF). Both astrocytes and neurons exhibit brilliant fluorochromasia within minutes of CFDA loading. However, following a brief rinse in buffered saline in the absence of CFDA, the astrocytes rapidly lose their cellular fluorescence while the neurons retain the dye for several hours. The fluorochromasia is uniformly distributed throughout the soma and processes which greatly facilitates the morphological identification of viable neurons. In addition, this protocol can be used to conveniently quantify neuronal survival in assays of the activities of neurotrophic or neurotoxic substances.

引用

页码：23 / 32

页数：10

共 36 条

[11] THE EXPRESSION AND DISTRIBUTION OF THE MICROTUBULE-ASSOCIATED PROTEINS-TAU AND MICROTUBULE-ASSOCIATED PROTEIN-2 IN HIPPOCAMPAL-NEURONS IN THE RAT INSITU AND IN CELL-CULTURE [J].