A direct microfluorometric method for measuring subpicomole amounts of nicotinamide adenine dinucleotide phosphate, glucose, and glycogen.
A direct microfluorometric method for measuring subpicomole amounts of nicotinamide adenine dinucleotide phosphate, glucose, and glycogen.
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DOI：
10.1006/abio.1998.2634
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发表时间：
1998-06
影响因子：
2.9
通讯作者：
L. T. Nguyen;D. Stephenson;G. Stephenson
中科院分区：
文献类型：
--
作者：
L. T. Nguyen;D. Stephenson;G. Stephenson
材料与方法显微荧光系统。在暗室中，用配备有双碱光电倍增管（Thorn EMI）和图表记录器（Activon）的倒置落射荧光显微镜（Axiovert 100 TV，Zeiss）测量荧光信号。使用氙灯（XBO 75 W）作为光源。蔡司滤光片系统包括激发滤光片（G 365）、分束器（FT 425）和发射滤光片（LP 420）。通过毛细管作用将荧光样品引入横截面为0.05 ± 0.5mm的矩形硼硅酸盐玻璃微池（Vitro Dynamics Inc.，新泽西州），这是放置在一个定制设计的黄铜框架较宽的一面。在15-20秒内记录当照射微池的1-mm部分（相当于25 nl）时产生的荧光信号。在测量过程中没有发生明显的漂白。
Materials and MethodsMicrofluorometric system. Fluorescence signals were measured, in a dark room, with an inverted epifluorescence microscope (Axiovert 100TV, Zeiss) equipped with a bialkali photomultiplier (Thorn EMI) and a chart recorder (Activon). A xenon lamp (XBO 75 W) was used as the light source. The Zeiss filter system included the excitation filter (G 365), the beamsplitter (FT 425), and the emission filter (LP 420). Fluorogenic samples were introduced by capillarity in rectangular borosilicate glass microcells with a cross section of 0.05 0.5 mm (Vitro Dynamics Inc., NJ), which were placed on the wider side on a custom-designed brass frame. Fluorescence signals generated when a 1-mm section of the microcell (equivalent to 25 nl) was illuminated were recorded over 15–20 s. No significant bleaching occurred during measurements.