Rotational dynamics of colloidal spheresprobed with fluorescence recovery after photobleaching

Lettinga, M. P.; Kuipers, B. W. M.; Koenderink, G. H.; Philipse, R. H.; Bessels, E.

doi:10.1063/1.1644799

Journal Article

Rotational dynamics of colloidal spheresprobed with fluorescence recovery after photobleaching

Lettinga, M. P.FZJ* ; Koenderink, G. H. ; Kuipers, B. W. M. ; Bessels, E. ; Philipse, R. H.

2004
American Institute of Physics Melville, NY

The journal of chemical physics 120, 4517 - 4529 (2004) [10.1063/1.1644799]

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Please use a persistent id in citations: http://hdl.handle.net/2128/1186 doi:10.1063/1.1644799

Abstract: We report a polarized fluorescence recovery after photobleaching (pFRAP) method to measure the rotational dynamics of fluorescent colloids over a wide dynamic range. The method is based on the polarization anisotropy in the fluorescence intensity, generated by bleaching of fluorescently labeled particles with an intense pulse of linearly polarized laser light. The rotational mobilities of the fluorescent particles can be extracted from the relaxation kinetics of the postbleach fluorescence polarization anisotropy. Our pFRAP setup has access to correlation times over a range of time scales from tens of microseconds to tens of seconds, and is highly sensitive, so very low concentrations of labeled particles can be probed. We present a detailed description of the theoretical background of pFRAP. The performance of the equipment is demonstrated for fluorescent colloidal silica spheres, dispersed in pure solvents as well as in fd-virus suspensions.

Keyword(s): Biomechanics (MeSH) ; Colloids: analysis (MeSH) ; Colloids: chemistry (MeSH) ; Computer Simulation (MeSH) ; Diffusion (MeSH) ; Fluorescence Recovery After Photobleaching: methods (MeSH) ; Microspheres (MeSH) ; Models, Chemical (MeSH) ; Models, Molecular (MeSH) ; Molecular Conformation (MeSH) ; Rotation (MeSH) ; Stress, Mechanical (MeSH) ; Colloids ; J