Protocol

[Overview] [Experimental Setup] [Protocol] [Data Evaluation] [Examples] [Literature]

[Preconditions] [Acquisition Bleaching] [Imaging Steps] [Prebleach] [Bleach] [Postbleach]

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There is no universal protocol for FRAP experiments since the design of a FRAP experiment always has to take into account the bleaching and recovery characteristics of the molecule under investigation. The visualized cells should not be affected by high illumination conditions.

Therefore the optimal conditions have to be empirically tested for each biological system.

The information in this section intends to provide general rules and hints applicable to most photobleaching experiments.

One general consideration in FRAP experiments is to minimize the bleaching during acquisition instead of acquiring “nice” images. The data has to be averaged over the selected area anyway to diminish statistical distributed noise.

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contact: Stefan Terjung           last update: 02/06/04