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Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics


  • Editor: Gregory W. Faris
  • Vol. 1, Iss. 6 — Jun. 13, 2006

Resolution enhancement in a light-sheet-based microscope (SPIM)

Christoph J. Engelbrecht and Ernst H.K. Stelzer  »View Author Affiliations

Optics Letters, Vol. 31, Issue 10, pp. 1477-1479 (2006)

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Light-sheet-based microscopy [single-plane illumination microscope (SPIM)] performs very well at low numerical apertures. It complements conventional (FM), confocal (CFM), and two-photon fluorescence microscopy ( 2 h ν -FM) currently used in modern life sciences. Lateral and axial SPIM point spread function (PSF) extents are measured by using fluorescent beads to determine the 3D resolution. The results are compared with values derived from an analytical theory and numerical simulations. The discrepancies are found to be less than 5%. The axial extent of a SPIM–PSF ( 10 × 0.3 W ) is approximately 5.7 μ m . This value is almost a factor of 2 smaller than in CFM, more than 2.5 times smaller than in FM, and more than three times smaller than in 2 h ν -FM. SPIM outperforms 2 h ν -FM and FM, while CFM has a better axial resolution at NAs above 0.8.

© 2006 Optical Society of America

OCIS Codes
(070.2580) Fourier optics and signal processing : Paraxial wave optics
(170.2520) Medical optics and biotechnology : Fluorescence microscopy
(170.6900) Medical optics and biotechnology : Three-dimensional microscopy
(180.2520) Microscopy : Fluorescence microscopy
(180.6900) Microscopy : Three-dimensional microscopy

ToC Category:
Medical Optics and Biotechnology

Original Manuscript: September 27, 2005
Revised Manuscript: January 5, 2006
Manuscript Accepted: January 17, 2006

Virtual Issues
Vol. 1, Iss. 6 Virtual Journal for Biomedical Optics

Christoph J. Engelbrecht and Ernst H. Stelzer, "Resolution enhancement in a light-sheet-based microscope (SPIM)," Opt. Lett. 31, 1477-1479 (2006)

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