In biological research, fluorescent proteins (FPs) are widely used in fluorescence microscopy. FPs are genetically encoded fluorophores, which are produced by cells after a noninvasive transfection. These proteins are utilized as markers and bio-sensors to study biological processes in living cells at high spatial and temporal resolution. Currently, many different types of FPs exist, comprising different spectral properties (i.e. colors), fluorescence lifetime characteristics, photo switching behavior, brightness, pH sensitivity, etc.
We developed a multi-position fluorescence lifetime imaging (FLIM) screening method to screen for bright FPs. However, this method can be applied to any experiment in which the fluorescence lifetime is an important parameter. To be able to test many different FP mutants (samples), an efficient acquisition and data processing workflow is paramount to obtain high quality and reproducible data with high throughput. The LIFA system, with the aid of MATLAB® and ImageJ , can be utilized to set up such an automated screening and characterization approach, allowing full flexibility.
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