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A Surface-Coupled Optical Trap with 1-bp Precision via Active Stabilization.

Stephen R Okoniewski^1,2, Ashley R Carter³, Thomas T Perkins^4,5

¹JILA, National Institute of Standards and Technology, and University of Colorado, Boulder, CO, 80309, USA.

Methods in Molecular Biology (Clifton, N.J.)

|November 16, 2016

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Force-Activated DNA Substrates for In Situ Generation of ssDNA and Designed ssDNA/dsDNA Structures in an Optical-Trapping Assay.

Methods in molecular biology (Clifton, N.J.)·2022

Improving optical trap stability is key for precise molecular motor studies. Active feedback loops minimize laser and sample motion, enabling Å-scale stability for 1-bp resolution.

Area of Science:

Biophysics
Optical Trapping
Molecular Motors

Background:

Optical traps offer Å-scale precision for bead motion measurement.
Inferring 1-base pair (bp) DNA motion of molecular motors is challenging due to noise sources degrading instrumental stability.

Purpose of the Study:

To detail methods for enhancing instrumental stability in optical traps.
To achieve Å-scale precision for single-base pair resolution in molecular motor studies.

Main Methods:

Minimizing laser noise (pointing, mode, polarization, intensity) using acousto-optical-modulator feedback.
Minimizing sample motion via a three-axis piezo-electric stage feedback loop.
Implementing active stabilization techniques for optical trapping.

Keywords:

Active stabilization Force spectroscopy Optical trap Optical tweezers Single molecule

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Main Results:

Achieved surface stability of 1 Å in 3D over tens of seconds.
Demonstrated 1-bp stability and precision in a surface-coupled optical trap over a broad bandwidth (0.03-2 Hz) at low force (6 pN).

Conclusions:

Active stabilization techniques are critical for achieving high precision in optical trap measurements.
These methods enhance laser and sample stability, benefiting biophysical assays like atomic force microscopy and super-resolution imaging.