A modular framework for post-processing and analysis of fluorescence microscopy image sequences of subcellular calcium dynamics

Abstract

Calcium (Ca2+) signaling is essential for activation of Tlymphocytes and can be understood as fundamental on-switch for the adaptive immune system. The activation is supposed to start by initial spatially and temporally localized Ca2+ signals. Imaging and analysis of these signals require high spatio-temporal resolution fluorescence microscopy – which, in turn, results in the need for an efficient and reliable post-processing and analysis workflow of the acquired image data. Started with a well established but time-consuming post-processing process, we report on our efforts to automatize and optimize it. The efforts led to a modular post-processing and analysis framework, which is presented. In addition, the influence of instances of the main blocks of the framework (e.g. bleaching correction, deconvolution) on Ca2+ dynamics analysis measures is evaluated.