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Updated: Aug 24, 2026
Measurement of Calcium Fluctuations Within the Sarcoplasmic Reticulum of Cultured Smooth Muscle Cells Using FRET-based Confocal Imaging
Published on: June 20, 2016
[Fluorescence quantification of intracellular calcium of rat smooth muscle cells]
Shui-yun Wang1, Fang Yan, He-xin Peng
1Department of Emergency, Shenzhen People's Hospital, Shenzhen 518020, China.
Objective:
To explore the kinetic changes of calcium in rat smooth muscle cells and establish a method for quantification of intracellular calcium.
Methods:
Rat mesenteric arteriolar smooth muscle cells (ASMCs) were isolated and the kinetic changes of calcium were measured using highly sensitive Ca2+ fluorescent probe indo-1 with laser scanning confocal microscopy (LSCM). The dissociation constant values (Kd) of the fluorescent probe indo-1 was measured at 37 degrees Celsius, and according to the conversion formula from fluorescence intensity to concentration, the concentration of Ca2+ was calculated.
Results:
Analysis of the fluorescent images using LSCM showed that [Ca2+]i in the ASMCs were significantly elevated in response to stimulation with dexamethasone, and spontaneous calcium waves as well as intracellular calcium overloading were observed occasionally.
Conclusion:
Fluorescence quantification with LSCM is applicable for detecting the kinetic changes of intracellular [Ca2+]i.
