OSA's Digital Library

Biomedical Optics Express

Biomedical Optics Express

  • Editor: Joseph A. Izatt
  • Vol. 3, Iss. 1 — Jan. 1, 2012
  • pp: 206–214

Improving signal-to-noise ratio of structured light microscopy based on photon reassignment

Vijay Raj Singh, Heejin Choi, Elijah Y. S. Yew, Dipanjan Bhattacharya, Luo Yuan, Colin J. R. Sheppard, Jagath C. Rajapakse, George Barbastathis, and Peter T. C. So  »View Author Affiliations

Biomedical Optics Express, Vol. 3, Issue 1, pp. 206-214 (2012)

View Full Text Article

Enhanced HTML    Acrobat PDF (889 KB)

Browse Journals / Lookup Meetings

Browse by Journal and Year


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools



In this paper, we report a method for 3D visualization of a biological specimen utilizing a structured light wide-field microscopic imaging system. This method improves on existing structured light imaging modalities by reassigning fluorescence photons generated from off-focal plane excitation, improving in-focus signal strength. Utilizing a maximum likelihood approach, we identify the most likely fluorophore distribution in 3D that will produce the observed image stacks under structured and uniform illumination using an iterative maximization algorithm. Our results show the optical sectioning capability of tissue specimens while mostly preserving image stack photon count, which is usually not achievable with other existing structured light imaging methods.

© 2011 OSA

OCIS Codes
(100.3020) Image processing : Image reconstruction-restoration
(110.6880) Imaging systems : Three-dimensional image acquisition
(180.2520) Microscopy : Fluorescence microscopy
(180.6900) Microscopy : Three-dimensional microscopy

ToC Category:

Original Manuscript: August 16, 2011
Revised Manuscript: December 11, 2011
Manuscript Accepted: December 11, 2011
Published: December 22, 2011

Virtual Issues
Advances in Optics for Biotechnology, Medicine, and Surgery (2011) Biomedical Optics Express

Vijay Raj Singh, Heejin Choi, Elijah Y. S. Yew, Dipanjan Bhattacharya, Luo Yuan, Colin J. R. Sheppard, Jagath C. Rajapakse, George Barbastathis, and Peter T. C. So, "Improving signal-to-noise ratio of structured light microscopy based on photon reassignment," Biomed. Opt. Express 3, 206-214 (2012)

Sort:  Author  |  Year  |  Journal  |  Reset  


  1. D. A. Agard, Y. Hiraoka, P. Shaw, and J. W. Sedat, “Fluorescence microscopy in three dimensions,” Methods Cell Biol.30, 353–377 (1989). [CrossRef] [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. D. Karadaglić and T. Wilson, “Image formation in structured illumination wide-field fluorescence microscopy,” Micron39(7), 808–818 (2008). [CrossRef] [PubMed]
  4. M. G. Gustafsson, L. Shao, P. M. Carlton, C. J. R. Wang, I. N. Golubovskaya, W. Z. Cande, D. A. Agard, and J. W. Sedat, “Three-dimensional resolution doubling in wide-field fluorescence microscopy by structured illumination,” Biophys. J.94(12), 4957–4970 (2008). [CrossRef] [PubMed]
  5. D. Lim, K. K. Chu, and J. Mertz, “Wide-field fluorescence sectioning with hybrid speckle and uniform-illumination microscopy,” Opt. Lett.33(16), 1819–1821 (2008). [CrossRef] [PubMed]
  6. 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]
  7. J. W. Goodman, Introduction to Fourier Optics (McGraw-Hill, 1996).
  8. M. Gu, Principles of Three-Dimensional Imaging in Confocal Microscopes (World Scientific, 1996), Chap. 5.
  9. D. L. Snyder, Random Point Processes (Wiley, New York, 1975), Chap. 6.
  10. A.van den Bos, Parameter Estimation for Scientists and Engineers (Wiley-Interscience, 2007).
  11. B. Richards and E. Wolf, “Electromagnetic diffraction in optical systems. II. Structure of the image field in an aplanatic system,” Proc. R. Soc. Lond. A Math. Phys. Sci.253(1274), 358–379 (1959). [CrossRef]
  12. V. Krishnamurthi, Y.-H. Liu, S. Bhattacharyya, J. N. Turner, and T. J. Holmes, “Blind deconvolution of fluorescence micrographs by maximum-likelihood estimation,” Appl. Opt.34(29), 6633–6647 (1995). [CrossRef] [PubMed]

Cited By

Alert me when this paper is cited

OSA is able to provide readers links to articles that cite this paper by participating in CrossRef's Cited-By Linking service. CrossRef includes content from more than 3000 publishers and societies. In addition to listing OSA journal articles that cite this paper, citing articles from other participating publishers will also be listed.


Fig. 1 Fig. 2 Fig. 3
Fig. 4 Fig. 5

« Previous Article

OSA is a member of CrossRef.

CrossCheck Deposited