Ratiometric Ca+ measurements using the FlexStation Scanning Fluorometer.
Ratiometric Ca+ measurements using the FlexStation Scanning Fluorometer.
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DOI：
10.1385/1-59259-949-4:119
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发表时间：
2005-01-01
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影响因子：
--
通讯作者：
McNulty, Shaun
中科院分区：
文献类型：
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作者：
Marshall, Ian C B;Boyfield, Izzy;McNulty, Shaun
目前，许多商业机构使用荧光成像酶标仪（FLIPR：Molecular Devices，桑尼维尔，CA）对细胞内Ca（2+）浓度进行高通量测量（见第7章），利用其快速动力学、可靠性和自动化兼容性。对于大多数工业应用，FLIPR的主要限制（即，其对使用可见光激发的单波长荧光探针的要求）不是重要问题。事实上，可见光探针提供了某些优于其紫外（UV）激发的比率测量对应物的益处，例如减少的样品自发荧光和更高的吸光度，从而允许使用相对低浓度的染料。然而，在某些情况下，研究人员可能更喜欢使用比率染料进行高通量实验，特别是当染料泄漏，光漂白或信噪比成为问题时。
Many commercial organizations currently use the Fluorometric Imaging Plate Reader (FLIPR: Molecular Devices, Sunnyvale, CA) to conduct high-throughput measurements of intracellular Ca(2+) concentration (see Chapter 7 ), taking advantage of its rapid kinetics, reliability, and compatibility for automation. For the majority of industrial applications, the primary limitation of FLIPR (i.e., its requirement for single wavelength fluorescent probes using visible light excitation) is not a significant issue. Indeed, visible light probes offer certain benefits over their ultraviolet (UV)-excited ratiometric counterparts, such as reduced sample autofluorescence and higher absorbance, thereby allowing relatively low concentrations of dye to be used. However, under certain circumstances researchers may prefer to conduct high-throughput experiments with ratiometric dyes, particularly when issues of dye leakage, photobleaching, or signal-to-noise ratio become a concern.