Fluorophores

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[How FRAP works] [Photobleaching] [Molecule Diffusion] [Recovery Dynamics] [Fluorophores]

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The optimal fluorophore should be bright and stable under low intensity illumination during image acquisition (pre- and postbleach). Under high intensity illumination it should bleach fast and irriversibly. Another prerequisite is that the used fluorophore must not inhibit the function of the investigated molecule.

Fluorophores for FRAP-experiments (according to Ellenberg and Rabut):

fluorophore

comment

fluorescein and derivatives

probably the most used chemical fluorophore, tends to bleach too easily and photoleaching is partly reversible (Periasamy 1996)

fluorescent proteins

mostly irreversible photobleaching
EGFP tends to multimerize  at high concentrations

Regardless of fluorophore, it is important to test itīs photobleaching characteristics under the imaging condition used.

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