A RAPID FLUOROMETRIC ASSAY TO MEASURE NEURONAL SURVIVAL IN-VITRO
A RAPID FLUOROMETRIC ASSAY TO MEASURE NEURONAL SURVIVAL IN-VITRO
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DOI：
10.1016/0165-0270(93)90009-g
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发表时间：
1993-11-01
影响因子：
3
通讯作者：
NEFF, NT
中科院分区：
文献类型：
--
作者：
BOZYCZKOCOYNE, D;MCKENNA, BW;NEFF, NT
我们报告的发展和特点的快速荧光微量测定适合定量神经元细胞存活。该方法可以以两种形式使用：（1）生存反应的时程分析或（2）作为评估各种试剂促进的神经元存活的简单终点测定。该检测使用钙黄绿素AM，一种非荧光、电中性、非极性的荧光素二乙酸酯类似物，其被动穿过细胞膜并被非特异性细胞内酯酶裂解为荧光衍生物。一旦在活细胞中裂解，所得荧光盐被完整的细胞膜保留。各种条件下活细胞的相对数量可以通过测量发射的荧光来定量。本文描述了允许测定低活神经元细胞数（10（2）-10（3）个细胞/cm（2））的条件。
We report the development and characterization of a rapid fluorometric microassay suitable for quantifying neuronal cell survival. The method can be used in two formats: (1) a time course analysis of survival response or (2) as a simple endpoint assay for the assessment of neuronal survival promoted by a variety of reagents. The assay uses calcein AM, a non-fluorescent, electrically neutral, non-polar analogue of fluorescein diacetate, which passively crosses cell membranes and is cleaved to a fluorescent derivative by non-specific intracellular esterases. Once cleaved in viable cells, the resultant fluorescent salts are retained by intact cell membranes. The relative number of viable cells under various conditions can be quantified by measuring the emitted fluorescence. Described herein are the conditions that allow the determination of low viable neuronal cell numbers (10(2)-10(3) cells/cm(2)).