scispace - formally typeset
Journal ArticleDOI

Stabilization of an optical microscope to 0.1 nm in three dimensions

Reads0
Chats0
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
More filters
Journal ArticleDOI

Single-molecule force spectroscopy: optical tweezers, magnetic tweezers and atomic force microscopy

TL;DR: These techniques are described and illustrated with examples highlighting current capabilities and limitations of single-molecule force spectroscopy.
Journal ArticleDOI

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.
Journal ArticleDOI

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

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
More filters
Journal ArticleDOI

Myosin V Walks Hand-Over-Hand: Single Fluorophore Imaging with 1.5-nm Localization

TL;DR: The results strongly support a hand-over-hand model of motility, not an inchworm model, which moves processively on actin.
Journal ArticleDOI

Direct observation of kinesin stepping by optical trapping interferometry

TL;DR: It is found that kinesin moves with 8-nm steps, similar to biological motors that move with regular steps.
Journal ArticleDOI

Single myosin molecule mechanics: piconewton forces and nanometre steps

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.
Journal ArticleDOI

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

TL;DR: The results suggest that kinesin is bound by both heads to the microtubule while it waits for adenosine triphosphate in between steps, and strongly support a hand-over-hand mechanism, and not an inchworm mechanism.
Related Papers (5)