Single-Image Diffusion Coefficient Measurements of Proteins in Free Solution
| Autor: | Je-Luen Li, Jonathan Kessler, Shannon Kian Zareh, Michael C. DeSantis, Yan Mei Wang |
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| Rok vydání: | 2012 |
| Předmět: |
Time Factors
Green Fluorescent Proteins Kinetics Spectroscopy Imaging and Other Techniques Analytical chemistry Biophysics FOS: Physical sciences Condensed Matter - Soft Condensed Matter Quantitative Biology - Quantitative Methods Diffusion Physics - Biological Physics Particle Size Diffusion (business) Quantitative Methods (q-bio.QM) Range (particle radiation) Chemistry Biomolecules (q-bio.BM) Fluorescence Molecular Imaging Computational physics Solutions Quantitative Biology - Biomolecules Biological Physics (physics.bio-ph) Temporal resolution FOS: Biological sciences Solvents Particle Soft Condensed Matter (cond-mat.soft) Particle size Intensity (heat transfer) |
| Zdroj: | Biophysical Journal. 102(7):1685-1691 |
| ISSN: | 0006-3495 |
| DOI: | 10.1016/j.bpj.2012.02.030 |
| Popis: | Diffusion coefficient measurements are important for many biological and material investigations, such as studies of particle dynamics and kinetics, and size determinations. Among current measurement methods, single particle tracking (SPT) offers the unique ability to simultaneously obtain location and diffusion information about a molecule while using only femtomoles of sample. However, the temporal resolution of SPT is limited to seconds for single-color-labeled samples. By directly imaging three-dimensional diffusing fluorescent proteins and studying the widths of their intensity profiles, we were able to determine the proteins' diffusion coefficients using single protein images of submillisecond exposure times. This simple method improves the temporal resolution of diffusion coefficient measurements to submilliseconds, and can be readily applied to a range of particle sizes in SPT investigations and applications in which diffusion coefficient measurements are needed, such as reaction kinetics and particle size determinations. |
| Databáze: | OpenAIRE |
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