Workflow and Methods of High-Content Time-Lapse Analysis for Quantifying Intracellular Calcium Signals

Abstract

Calcium ions (Ca 2+ ) play a fundamental role in a variety of physiological functions in many cell types by acting as a secondary messenger. Variation of intracellular Ca 2+ concentration ([Ca 2+ ] i ) is often observed when the cell is stimulated. However, it is a challenging task to automatically quantify intracellular [Ca 2+ ] 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 H 2 O 2 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

Tracking transcriptional activities with high-content epifluorescent imaging

“…These fusion-reporters can be used in large scale RNAi or small molecules screening, in time-lapse study and/or case-control study. [7][8][9][10][11] However, to serve as phenotype indicators, only a few key reporters are needed, often less than three. Hence such approaches provide little insights on the mechanism of the cellular regulation network in response to external perturbations.…”