[Fluorescence quantification of intracellular calcium of rat smooth muscle cells].
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.


CONCLUSIONS
Fluorescence quantification with LSCM is applicable for detecting the kinetic changes of intracellular [Ca2+]i.
Wang
Shui-yun
SY
Department of Emergency, Shenzhen People's Hospital, Shenzhen 518020, China.
Yan
Fang
F
Peng
He-xin
HX
Zheng
Chang-dong
CD
Bai
Cheng-wen
CW
D000818:Animals; D002118:Calcium / Q000032:analysis*; D003907:Dexamethasone / Q000494:pharmacology; D005260:Female; D005453:Fluorescence; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000737:chemistry* / Q000166:cytology; D032389:Myocytes, Smooth Muscle / Q000737:chemistry*; D051381:Rats; D017207:Rats, Sprague-Dawley
D003907:Dexamethasone; D002118:Calcium