Workflow and methods of high-content time-lapse analysis for quantifying intracellular calcium signals.
Calcium ions (Ca2+) play a fundamental role in a variety of physiological functions in many cell types by acting as a secondary messenger. Variation of intracellular Ca2+ concentration ([Ca2+]i) is often observed when the cell is stimulated. However, it is a challenging task to automatically quantify intracellular [Ca2+]i in a population of cells. In this study, we present a workflow including specific algorithms for the automated intracellular calcium signal analysis using high-content, time-lapse cellular images. The experimental validations indicate the effectiveness of the proposed workflow and algorithms. We applied the workflow to analyze the intracellular calcium signals induced by different concentrations of H2O2 in the cell lines transfected by presenilin-1 (PS-1) that is known to be closely related to the familial Alzheimer's disease (FAD). The analysis results imply an important role of mutant PS-1, but not normal human PS-1 and mutant human amyloid precursor protein (APP), in enhancing intracellular calcium signaling induced by H2O2.
Department of Information Science, School of Mathematical Sciences, and LMAM, Peking University, Beijing, 100871, China.
Fuhai
F
Li
Xiaobo
X
Zhou
Jinmin
J
Zhu
Weiming
W
Xia
Jinwen
J
Ma
Stephen T C
ST
Wong
D000465:Algorithms; D000544:Alzheimer Disease / Q000235:genetics / Q000378:metabolism / Q000503:physiopathology; D000818:Animals; D016466:CHO Cells; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling / Q000502:physiology*; D006224:Cricetinae; D003412:Cricetulus; D001330:Electronic Data Processing / Q000379:methods; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007424:Intracellular Fluid / Q000737:chemistry / Q000378:metabolism*; D018384:Oxidative Stress / Q000187:drug effects / Q000235:genetics; D053764:Presenilin-1 / Q000235:genetics / Q000378:metabolism; D012815:Signal Processing, Computer-Assisted; D012984:Software; D012986:Software Validation; D013997:Time Factors
D053764:Presenilin-1; D006861:Hydrogen Peroxide; D002118:Calcium