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

  • Stephen R. Okoniewski
  • Ashley R. Carter
  • Thomas T. Perkins
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1486)

Abstract

Optical traps can measure bead motions with Å-scale precision. However, using this level of precision to infer 1-bp motion of molecular motors along DNA is difficult, since a variety of noise sources degrade instrumental stability. In this chapter, we detail how to improve instrumental stability by (1) minimizing laser pointing, mode, polarization, and intensity noise using an acousto-optical-modulator mediated feedback loop and (2) minimizing sample motion relative to the optical trap using a three-axis piezo-electric-stage mediated feedback loop. These active techniques play a critical role in achieving a surface stability of 1 Å in 3D over tens of seconds and a 1-bp stability and precision in a surface-coupled optical trap over a broad bandwidth (Δf = 0.03–2 Hz) at low force (6 pN). These active stabilization techniques can also aid other biophysical assays that would benefit from improved laser stability and/or Å-scale sample stability, such as atomic force microscopy and super-resolution imaging.

Key words

Optical trap Optical tweezers Single molecule Active stabilization Force spectroscopy 

Notes

Acknowledgments

We thank Carl Sauer for providing detailed electronic diagrams and associated files for the circuit boards. This work is supported by a National Science Foundation Graduate Research Fellowship (Grant No. DGE 1144083 to S.R.O.), a National Institute of Health Molecular Biophysics Training Grant awarded to S.R.O. (T32 GM-065103), the NSF (Phys-1125844), and NIST. Mention of commercial products is for information only; it does not imply NIST recommendation or endorsement, nor does it imply that the products mentioned are necessarily the best available for the purpose. T.T.P. is a staff member of NIST’s quantum physics division.

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Stephen R. Okoniewski
    • 1
    • 2
  • Ashley R. Carter
    • 3
  • Thomas T. Perkins
    • 1
    • 4
  1. 1.JILANational Institute of Standards and Technology, and University of ColoradoBoulderUSA
  2. 2.Department of PhysicsUniversity of ColoradoBoulderUSA
  3. 3.Department of Physics and AstronomyAmherst CollegeAmherstUSA
  4. 4.Department of Molecular, Cellular, and Developmental BiologyUniversity of ColoradoBoulderUSA

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