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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)
http://dx.doi.org/10.1364/BOE.3.000170


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Abstract

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:
Microscopy

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

Citation
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)
http://www.opticsinfobase.org/boe/abstract.cfm?URI=boe-3-1-170


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References

  1. P. A. Santi, S. B. Johnson, M. Hillenbrand, P. Z. GrandPre, T. J. Glass, and J. R. Leger, “Thin-sheet laser imaging microscopy for optical sectioning of thick tissues,” Biotechniques46(4), 287–294 (2009). [PubMed]
  2. M. A. A. Neil, R. Juskaitis, and T. Wilson, “Method of obtaining optical sectioning by using structured light in a conventional microscope,” Opt. Lett.22(24), 1905–1907 (1997). [CrossRef] [PubMed]
  3. J. Mertz and J. Kim, “Scanning light-sheet microscopy in the whole mouse brain with HiLo background rejection,” J. Biomed. Opt.15(1), 016027 (2010). [CrossRef] [PubMed]
  4. P. J. Keller and E. H. Stelzer, “Quantitative in vivo imaging of entire embryos with digital scanned laser light sheet fluorescence microscopy,” Curr. Opin. Neurobiol.18(6), 624–632 (2008). [CrossRef] [PubMed]
  5. P. J. Keller, A. D. Schmidt, A. Santella, K. Khairy, Z. Bao, J. Wittbrodt, and E. H. Stelzer, “Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy,” Nat. Methods7(8), 637–642 (2010). [CrossRef] [PubMed]
  6. P. Schacht, S. B. Johnson, and P. A. Santi, “Implementation of a continuous scanning procedure and a line scan camera for thin-sheet laser imaging microscopy,” Biomed. Opt. Express1(2), 598–609 (2010). [CrossRef] [PubMed]
  7. J. A. N. Buytaert and J. J. J. Dirckx, “Design and quantitative resolution measurements of an optical virtual sectioning three-dimensional imaging technique for biomedical specimens, featuring two-micrometer slicing resolution,” J. Biomed. Opt.12(1), 014039 (2007). [CrossRef] [PubMed]
  8. L. H. Schaefer, D. Schuster, and J. Schaffer, “Structured illumination microscopy: artefact analysis and reduction utilizing a parameter optimization approach,” J. Microsc.216(2), 165–174 (2004). [CrossRef] [PubMed]

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