An algorithm for automated detection, localization and measurement of local calcium signals from camera-based imaging

作者：Ellefsen Kyle L; Settle Brett; Parker Ian; Smith Ian F^*

来源：Cell Calcium, 2014, 56(3): 147-156.

DOI：10.1016/j.ceca.2014.06.003

• Calcium
• Imaging
• Total internal reflection microscopy
• Algorithm
• Automation
• Fluorescence

Local Ca2+ transients such as puffs and sparks form the building blocks of cellular Ca2+ signaling in numerous cell types. They have traditionally been studied by linescan confocal microscopy, but advances in TIRF microscopy together with improved electron-multiplied CCD (EMCCD) cameras now enable rapid (>500 frames s(-1)) imaging of subcellular Ca2+ signals with high spatial resolution in two dimensions. This approach yields vastly more information (ca. 1 Gb min(-1)) than linescan imaging, rendering visual identification and analysis of local events imaged both laborious and subject to user bias. Here we describe a routine to rapidly automate identification and analysis of local Ca2+ events. This features an intuitive graphical user-interfaces and runs under Matlab and the open-source Python software. The underlying algorithm features spatial and temporal noise filtering to reliably detect even small events in the presence of noisy and fluctuating baselines; localizes sites of Ca2+ release with sub-pixel resolution; facilitates user review and editing of data; and outputs time-sequences of fluorescence ratio signals for identified event sites along with Excel-compatible tables listing amplitudes and kinetics of events.

• 出版日期2014-9