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Applied Optics

Applied Optics


  • Editor: James C. Wyant
  • Vol. 47, Iss. 12 — Apr. 20, 2008
  • pp: 2070–2075

Stabilizing method for reflection interference contrast microscopy

Kipom Kim and Omar A. Saleh  »View Author Affiliations

Applied Optics, Vol. 47, Issue 12, pp. 2070-2075 (2008)

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Reflection interference contrast microscopy (RICM) is a technique for measuring the shape and position of microscopic objects in solution; it has many biological and biophysical applications. Use of RICM for long-time acquisitions requires minimizing defocusing effects that are due to thermal and mechanical drift. We present a simple stabilizing method that accomplishes this using an image-analysis-based linear focus function to establish feedback control of the focal position. While implementing this routine, we used RICM for independent measurement of the apparent fluctuation in the vertical position of an immobilized bead: the measured height had a standard deviation of 0.12 nm during a 45 min acquisition while under feedback control, demonstrating the high stability achievable with our approach.

© 2008 Optical Society of America

OCIS Codes
(120.2650) Instrumentation, measurement, and metrology : Fringe analysis
(120.3180) Instrumentation, measurement, and metrology : Interferometry
(170.6900) Medical optics and biotechnology : Three-dimensional microscopy
(180.3170) Microscopy : Interference microscopy

ToC Category:
Instrumentation, Measurement, and Metrology

Original Manuscript: August 16, 2007
Revised Manuscript: February 19, 2008
Manuscript Accepted: February 19, 2008
Published: April 16, 2008

Virtual Issues
Vol. 3, Iss. 5 Virtual Journal for Biomedical Optics

Kipom Kim and Omar A. Saleh, "Stabilizing method for reflection interference contrast microscopy," Appl. Opt. 47, 2070-2075 (2008)

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