Particle tracking and TILLvisTRAC package

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Introduction

Modern fluorescence microscopy systems (like our Polychrome 5000−based imaging solution) in combination with specific fluorescent markers have been widely employed to study dynamic events in living cells e.g. membrane trafficking, cell motility or cytoskeleton dynamics. Such experiments generate large and complex data sets which are difficult to interpret in a consistent manner when analysed manually. It should also not be overlooked that such an approach is monotonous and very time consuming. In addition, the signal to noise ratio of the objects is often low and may vary through out the image sequence. This is true, for example, in experiments where small fast moving vesicles are to be observed − here the poor S/N ratio is caused by the short camera integration times used when capturing images rapidly. On the other hand slow dynamic processes, such as chromatin movements in the nucleus, require extended periods of observation − here poor S/N results because total light exposure must be kept at a minimum (hence the use of short camera integration times) to avoid phototoxic effects.

Our solution is a comprehensive software module − TILLvisTRAC. This incorporates a novel technique for automated analysis and time−space visualization with sub−pixel resolution of time series from living cells. The TILLvisTRAC software is based on the published method of Tvaruskó et al.