Automatic Detection and Classification of Ca2+ Release Events in Confocal Line- and Framescan Images

Abstractabstract text

Introduction

Numerous methods exist for analysing sparks in confocal linescans. These employ various different approaches: noise tresholding (Ríos 2001, Picht 2007), wavelet transform (Szabó 2010), etc.

Recently, a method was developed by Tian et al., (Tian 2012), where fluorescence time trace in each pixel is fitted and provides a practically noise free approximation of the original fluorescence data. This pixel-by-pixel method had, however, several limitations which made it impractical to be used for Ca2+ release event detection.

Here we extend the method by Tian et al., in several ways. The new method allows for Ca2+ release event classification based on pixel-by-pixel denoising of the original signal.

Methods

Cell isolations

Experimental

Data acquisition was performed on two confocal setups. Linescan images were obtained on a Olympus FluoView 1000 confocal microscope. Framescan images were recorded with a VTInfinity multi-beam confocal microscope recording 512x64 pixel images at 150Hz freqency.

Computational

The detection algorithm is presented in the Results section.