Merits of various calcium indicators

Abstract

Ca2+ is a vital element that is involved in the regulation of a wide range of living cell functions. This study has aimed to examine the efficacy of various calcium indicators and their ability to detect Ca2+ .The indicator Cal-520 AM (5µM) and Fluo-4 AM (5µM) were used to detect calcium ions as tested on endothelial tissue from the arteries of rats. Six experiments were applied, and six rats were used and divided into two groups of indicators. Arterial endothelial cells obtained by biopsy were used in the microscope step, transferred to an imaging chamber, and viewed in an inverted microscope. Analysis of endothelial Ca2+ signalling imaging was done using an inverted fluorescence microscope (Ti Eclipse Nikon, Japan). Then, the results of images were uploaded to ImageJ software to measure fluorescence (F) intensity. After adding acetylcholine (Ach) at 60 seconds, 600 samples were taken to study the results of reading of 600 time(s) to test the study hypothesis that is there a statistically significant difference between the Cal-520 and Fluo-4 indicators in detecting Ca2+ . A suitable analysis was chosen according to the variable type. Analyses were conducted using independent sample t tests, and sensitivity was conducted to measure the diagnostic test. The analysis showed that 100% of abnormal cases were classified by the test as positive. The statistical tests showed that there was a highly statistically significance difference between Cal-520 and Fluo-4. The efficacy trend was toward the Cal-520 indicator due to the mean average of the fluorescence intensity.