Journal ArticleDOI
Stabilization of an optical microscope to 0.1 nm in three dimensions
Ashley R. Carter,Gavin M. King,Theresa A. Ulrich,Wayne A. Halsey,David Alchenberger,Thomas T. Perkins +5 more
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TLDR
This work overcame mechanical drift and achieved atomic-scale stabilization (0.1 nm) of an optical microscope in 3D by measuring the position of a fiducial mark coupled to the microscope cover slip using back-focal-plane (BFP) detection and correcting for the drift using a piezoelectric stage.Abstract:
Mechanical drift is a long-standing problem in optical microscopy that occurs in all three dimensions. This drift increasingly limits the resolution of advanced surface-coupled, single-molecule experiments. We overcame this drift and achieved atomic-scale stabilization (0.1 nm) of an optical microscope in 3D. This was accomplished by measuring the position of a fiducial mark coupled to the microscope cover slip using back-focal-plane (BFP) detection and correcting for the drift using a piezoelectric stage. Several significant factors contributed to this experimental realization, including (i) dramatically reducing the low frequency noise in BFP detection, (ii) increasing the sensitivity of BFP detection to vertical motion, and (iii) fabricating a regular array of nanometer-sized fiducial marks that were firmly coupled to the cover slip. With these improvements, we achieved short-term (1 s) stabilities of 0.11, 0.10, and 0.09 nm (rms) and long-term (100 s) stabilities of 0.17, 0.12, and 0.35 nm (rms) in x, y, and z, respectively, as measured by an independent detection laser.read more
Citations
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Journal ArticleDOI
Single-molecule force spectroscopy: optical tweezers, magnetic tweezers and atomic force microscopy
Keir C. Neuman,Attila Nagy +1 more
TL;DR: These techniques are described and illustrated with examples highlighting current capabilities and limitations of single-molecule force spectroscopy.
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Recent Advances in Optical Tweezers
TL;DR: Although technical in nature, these developments have important implications for the expanded use of optical tweezers in biochemical research and thus should be of general interest.
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Optimized localization analysis for single-molecule tracking and super-resolution microscopy.
TL;DR: Both theory and experimental data showed that unweighted least-squares fitting of a Gaussian squanders one-third of the available information, a popular formula for its precision exaggerates beyond Fisher's information limit, and weighted least-Squares may do worse, whereas maximum-likelihood fitting is practically optimal.
Journal ArticleDOI
Optical tweezers study life under tension
Furqan M. Fazal,Steven M. Block +1 more
TL;DR: Optical tweezers have become one of the primary weapons in the arsenal of biophysicists, and have revolutionized the new field of single-molecule biophysics.
Journal ArticleDOI
Single-molecule biophysics: at the interface of biology, physics and chemistry
TL;DR: An overview of the burgeoning field of single-molecule biophysics is presented, discussing key highlights and selected examples from its genesis to the authors' projections for its future.
References
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Myosin V Walks Hand-Over-Hand: Single Fluorophore Imaging with 1.5-nm Localization
Ahmet Yildiz,Joseph N. Forkey,Sean A. McKinney,Taekjip Ha,Taekjip Ha,Yale E. Goldman,Paul R. Selvin,Paul R. Selvin +7 more
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TL;DR: A new in vitro assay using a feedback enhanced laser trap system allows direct measurement of force and displacement that results from the interaction of a single myosin molecule with a single suspended actin filament.
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Stretching DNA with optical tweezers
TL;DR: Force-extension (F-x) relationships were measured for single molecules of DNA under a variety of buffer conditions, using an optical trapping interferometer modified to incorporate feedback control, suggesting that the intrinsic persistence length remains close to 40 nm.
Journal ArticleDOI
Kinesin Walks Hand-Over-Hand
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