DOI: 10.1073/pnas.0403534101. Simultaneous atomic force microscope and fluorescence measurements of protein unfolding using a calibrated evanescent wave

Simultaneous atomic force microscope and fluorescence measurements of protein unfolding using a calibrated evanescent wave

10.1073/pnas.0403534101

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

Fluorescence techniques for monitoring single-molecule dynamics in the vertical dimension currently do not exist. Here we use an atomic force microscope to calibrate the distance-dependent intensity decay of an evanescent wave. The measured evanescent wave transfer function was then used to convert the vertical motions of a fluorescent particle into displacement (SD = <1 nm). We demonstrate the use of the calibrated evanescent wave to resolve the 20.1 ± 0.5-nm step increases in the length of the small protein ubiquitin during forced unfolding. The experiments that we report here make an important contribution to fluorescence microscopy by demonstrating the unambiguous optical tracking of a single molecule with a resolution comparable to that of an atomic force microscope.

Bibliography

Sarkar, A., Robertson, R. B., & Fernandez, J. M. (2004). Simultaneous atomic force microscope and fluorescence measurements of protein unfolding using a calibrated evanescent wave. Proceedings of the National Academy of Sciences, 101(35), 12882â12886.

Authors 3

Atom Sarkar (first)
Ragan B. Robertson (additional)
Julio M. Fernandez (additional)

References 23 Referenced 112

Dates

Type	When
Created	21 years ago (Aug. 23, 2004, 8:16 p.m.)
Deposited	3 years, 4 months ago (April 12, 2022, 9:30 a.m.)
Indexed	4 weeks, 1 day ago (July 28, 2025, 2:36 a.m.)
Issued	21 years ago (Aug. 23, 2004)
Published	21 years ago (Aug. 23, 2004)
Published Online	21 years ago (Aug. 23, 2004)
Published Print	20 years, 11 months ago (Aug. 31, 2004)

Funders 0

None

BibTeX

@article{Sarkar_2004, title={Simultaneous atomic force microscope and fluorescence measurements of protein unfolding using a calibrated evanescent wave}, volume={101}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.0403534101}, DOI={10.1073/pnas.0403534101}, number={35}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Sarkar, Atom and Robertson, Ragan B. and Fernandez, Julio M.}, year={2004}, month=aug, pages={12882–12886} }

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