不同光激发钙荧光探针的性质、选择标准及测定过程的标准化

Property,selection criteria and calibration of calcium fluorescent indicators

目的:概括钙荧光探针的分类,选择钙荧光探针的基本思路以及细胞内钙测定的标准化。资料来源:应用计算机检索Pubmed数据库2000-01/2006-12相关钙荧光探针方面的文献,检索词“calcium fluorescent indicators,calcium measurement”,限定文献语言种类为English。资料选择:对资料进行初审,选取包括钙荧光探针的文献,开始查找全文。纳入标准:与钙荧光探针及细胞内钙测定相关的文献。排除标准:重复研究、个案报告、综述、Meta分析的文献。资料提炼:共检索到103篇关于钙荧光探针及细胞内钙测定的文献,最终纳入30篇符合标准的文献。资料综合:钙荧光探针可分为定性探针及定量探针两类,定性探针多为可见光激发的单波长探针,其代表有Fluo3和Fluo4等,定量探针多为紫外激发的双波长探针,其代表为Fura2及Indo1等。选择钙荧光探针需考虑试验目的,探针进入细胞的方式,探针的离解常数及测定所用仪器等。测定细胞内钙时,应注重测定过程中的标准化,同时注意钙荧光探针测定过程中的潜在问题,避免混杂因素的干扰。本文主要综述如何选择钙荧光探针。结论:选择合适的钙探针,注意钙荧光探针测定过程中的标准化及其他潜在问题,可对细胞内钙离子浓度进行较为准确的测定。

OBJECTIVE： To explain the classification of calcium fluorescent indicators, introduce the selection cdteria of calcium fluorescent indicators and the calibration of intracellular calcium indicators. DATA SOURCES： Using the key terms of ＂calcium fluorescent indicators, calcium measurement＇, we searched the Medline database in English from January 2000 to December 2006 for the articles related to calcium fluorescent indicators. STUDY SELECTION： All articles were selected firstly, the articles relevant to calcium fluorescent indicators were selected and looked up for full texts. Inclusive criteria： the articles relevant to experiment research of the calcium fluorescent indicators and intracellular calcium measurement were excluded. Exclusive criteria： repeated research, case report, review or Meta analysis. DATA EXTRACTION： A total of 103 articles were searched. Finally we selected 30 articles that met the selection and exclusion criterion. DATA SYNTHESIS： The calcium fluorescent indicator is classified to two categories： the qualitative indictors and quantitative indictors. The typical qualitative indictors often refer to the single-wave indicators excited by visible lights and include Fluo 3 and Fluo 4 etc. The quantitative indictors often refer to the dual-wave indicators excited by ultraviolet lights and include Fura 2 and Indo 1 etc. To choose a proper calcium fluorescent indicator, we had to consider the indicator form, the way to reach the cellular environments, dissociation constant of indicator and the instruments that apply to measurement. In the practice of measuring intracellular calcium, we have to pay close attention to the calibration process and consider some potential problems, which may encounter in the calcium measurement. This paper is aimed to the selection of calcium fluorescent indictor. CONCLUSION： Selection of suitable calcium fluorescent indictor need close attention to the calibration process and consider some potential problems. We can measure the subtle change of intracellular calcium ion.