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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 20, Iss. 28 — Dec. 31, 2012
  • pp: 29353–29360

Autofocusing system based on optical astigmatism analysis of single-molecule images

Wonseok Hwang, Sangsu Bae, and Sungchul Hohng  »View Author Affiliations

Optics Express, Vol. 20, Issue 28, pp. 29353-29360 (2012)

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Single-molecule fluorescence imaging has greatly contributed to our understanding of many bio-molecular systems. While reactions occurring in the range of several minutes can be readily studied using conventional single-molecule fluorescence microscopes, data acquisition for longer time scales is hindered by the focal drift of high numerical aperture objectives, which should be corrected in real time. Here, we developed a robust autofocusing system based on optical astigmatism analysis of single-molecule images. Compared to the previously developed methods, our approach has a merit of simplicity in that neither fiducial makers nor an additional laser-detector system is required. As a demonstration, we observed B-Z transition dynamics occurring for several hours.

© 2012 OSA

OCIS Codes
(110.0180) Imaging systems : Microscopy
(180.2520) Microscopy : Fluorescence microscopy

ToC Category:

Original Manuscript: October 24, 2012
Revised Manuscript: November 29, 2012
Manuscript Accepted: December 2, 2012
Published: December 18, 2012

Virtual Issues
Vol. 8, Iss. 1 Virtual Journal for Biomedical Optics

Wonseok Hwang, Sangsu Bae, and Sungchul Hohng, "Autofocusing system based on optical astigmatism analysis of single-molecule images," Opt. Express 20, 29353-29360 (2012)

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