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Applied Optics

Applied Optics


  • Editor: Joseph N. Mait
  • Vol. 50, Iss. 25 — Sep. 1, 2011
  • pp: 4989–4997

Simple way of pinpointing the three-dimensional position of biomarkers in fluorescence microscopy using a through-focus exposure method

Koichiro Kishima  »View Author Affiliations

Applied Optics, Vol. 50, Issue 25, pp. 4989-4997 (2011)

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The author proposes a method to identify the three-dimensional positions of fluorescent biomarkers by recording just two images. In the proposed method, the x and y positions of all fluorescent markers are recorded in the first exposure, and the z positions are obtained from a blurred image in the second exposure. The author has verified this method using a specimen with 1 μm deep grooves and applied it to measuring chromatic aberration and the separation between two biological probes in fluorescence in situ hybridization cells. The method offers the advantage of greatly reduced data storage requirements.

© 2011 Optical Society of America

OCIS Codes
(110.4190) Imaging systems : Multiple imaging
(180.2520) Microscopy : Fluorescence microscopy
(180.6900) Microscopy : Three-dimensional microscopy

ToC Category:

Original Manuscript: March 28, 2011
Revised Manuscript: June 7, 2011
Manuscript Accepted: July 7, 2011
Published: August 29, 2011

Virtual Issues
Vol. 6, Iss. 10 Virtual Journal for Biomedical Optics

Koichiro Kishima, "Simple way of pinpointing the three-dimensional position of biomarkers in fluorescence microscopy using a through-focus exposure method," Appl. Opt. 50, 4989-4997 (2011)

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