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Biomedical Optics Express

Biomedical Optics Express

  • Editor: Joseph A. Izatt
  • Vol. 3, Iss. 1 — Jan. 1, 2012
  • pp: 170–177

Scanning thin-sheet laser imaging microscopy (sTSLIM) with structured illumination and HiLo background rejection.

Tobias J. Schröter, Shane B. Johnson, Kerstin John, and Peter A. Santi  »View Author Affiliations

Biomedical Optics Express, Vol. 3, Issue 1, pp. 170-177 (2012)

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We report replacement of one side of a static illumination, dual sided, thin-sheet laser imaging microscope (TSLIM) with an intensity modulated laser scanner in order to implement structured illumination (SI) and HiLo image demodulation techniques for background rejection. The new system is equipped with one static and one scanned light-sheet and is called a scanning thin-sheet laser imaging microscope (sTSLIM). It is an optimized version of a light-sheet fluorescent microscope that is designed to image large specimens (<15 mm in diameter). In this paper we describe the hardware and software modifications to TSLIM that allow for static and uniform light-sheet illumination with SI and HiLo image demodulation. The static light-sheet has a thickness of 3.2 µm; whereas, the scanned side has a light-sheet thickness of 4.2 µm. The scanned side images specimens with subcellular resolution (<1 µm lateral and <4 µm axial resolution) with a size up to 15 mm. SI and HiLo produce superior contrast compared to both the uniform static and scanned light-sheets. HiLo contrast was greater than SI and is faster and more robust than SI because as it produces images in two-thirds of the time and exhibits fewer intensity streaking artifacts.

© 2011 OSA

OCIS Codes
(170.3880) Medical optics and biotechnology : Medical and biological imaging
(180.0180) Microscopy : Microscopy
(180.2520) Microscopy : Fluorescence microscopy
(180.6900) Microscopy : Three-dimensional microscopy
(220.0220) Optical design and fabrication : Optical design and fabrication

ToC Category:

Original Manuscript: September 27, 2011
Revised Manuscript: December 8, 2011
Manuscript Accepted: December 15, 2011
Published: December 19, 2011

Tobias J. Schröter, Shane B. Johnson, Kerstin John, and Peter A. Santi, "Scanning thin-sheet laser imaging microscopy (sTSLIM) with structured illumination and HiLo background rejection.," Biomed. Opt. Express 3, 170-177 (2012)

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