High-resolution intracellular viscosity measurement using time-dependent fluorescence anisotropy
2010; Optica Publishing Group; Volume: 18; Issue: 16 Linguagem: Inglês
10.1364/oe.18.016607
ISSN1094-4087
AutoresWesley C. Parker, Nilay Chakraborty, Regina M. Vrikkis, Gloria D. Elliott, Stuart T. Smith, Patrick J. Moyer,
Tópico(s)Protein Interaction Studies and Fluorescence Analysis
ResumoA low-cost pulsed laser is used in conjunction with a homebuilt laser confocal-scanning epifluorescence microscope having submicron lateral and axial spatial resolution to determine cytoplasmic viscosity at specific intracytoplasmic locations in J774 mouse macrophage cells. Time-dependent fluorescence anisotropy measurements are made at each location and global deconvolution techniques are used to determine rotational correlation times. These rotational correlation times are related to the hydrated volume of 8-hydroxyperene-1,3,6-trisulfonic acid (HPTS) to calculate viscosity at specific points inside the cell. In the cytoplasmic areas measured, rotational correlation times of HPTS ranged from 0.186 ns to 0.411 ns, corresponding to viscosities ranging from 1.00 +/- 0.03 cP to 2.21+/- 0.05 cP.
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