Theoretical analysis on ratiometric fluorescent indicators caused biased estimates of intracellular free calcium concentrations |
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Authors: | Hu Zhiwen Yu Liangdeng Yu Zengliang |
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Institution: | Key Laboratory of Ion Beam Bioengineering, Institute of Plasma Physics, Chinese Academy of Sciences, P.O. Box 1126, Hefei, Anhui 230031, China. |
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Abstract: | Ratiometric fluorescent calcium indicator dyes have been widely used for the study of the role of Ca2+ in cell physiopathology. Although these ratiometric dyes offer several advantages over others, they suffer some drawbacks which cause serious errors in measurement of intracellular Ca2+ concentration, Ca2+]i. The present study systematically analyzes theoretical reasons and technical sources of discrepancies occurring in the measurement of the characteristics of the agonists-induced cells Ca2+]i. In order to avoid the errors and achieve the accurate determination of Ca2+]i, this study proposes solutions and suggests some critical measures in both theoretical and technical aspects. Therefore, this analysis can be a valuable tool in clarifying proper usages of fluorescent dyes for Ca2+]i measurements. |
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