OSA's Digital Library

Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 19, Iss. 15 — Jul. 18, 2011
  • pp: 14508–14517

Analyzing speckle contrast for HiLo microscopy optimization

J. Mazzaferri, D. Kunik, J. M. Belisle, K. Singh, S. Lefrançois, and S. Costantino  »View Author Affiliations

Optics Express, Vol. 19, Issue 15, pp. 14508-14517 (2011)

View Full Text Article

Enhanced HTML    Acrobat PDF (1332 KB)

Browse Journals / Lookup Meetings

Browse by Journal and Year


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools



HiLo microscopy is a recently developed technique that provides both optical sectioning and fast imaging with a simple implementation and at a very low cost. The methodology combines widefield and speckled illumination images to obtain one optically sectioned image. Hence, the characteristics of such speckle illumination ultimately determine the quality of HiLo images and the overall performance of the method. In this work, we study how speckle contrast influence local variations of fluorescence intensity and brightness profiles of thick samples. We present this article as a guide to adjust the parameters of the system for optimizing the capabilities of this novel technology.

© 2011 OSA

OCIS Codes
(110.6150) Imaging systems : Speckle imaging
(170.1790) Medical optics and biotechnology : Confocal microscopy
(170.6900) Medical optics and biotechnology : Three-dimensional microscopy
(180.2520) Microscopy : Fluorescence microscopy

ToC Category:

Original Manuscript: June 1, 2011
Revised Manuscript: June 30, 2011
Manuscript Accepted: July 5, 2011
Published: July 13, 2011

Virtual Issues
Vol. 6, Iss. 8 Virtual Journal for Biomedical Optics

J. Mazzaferri, D. Kunik, J. M. Belisle, K. Singh, S. Lefrançois, and S. Costantino, "Analyzing speckle contrast for HiLo microscopy optimization," Opt. Express 19, 14508-14517 (2011)

Sort:  Author  |  Year  |  Journal  |  Reset  


  1. 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]
  2. C. Ventalon, R. Heintzmann, and J. Mertz, “Dynamic speckle illumination microscopy with wavelet prefiltering,” Opt. Lett. 32(11), 1417–1419 (2007). [CrossRef] [PubMed]
  3. D. Lim, T. N. Ford, K. K. Chu, and J. Mertz, “Optically sectioned in vivo imaging with speckle illumination HiLo microscopy,” J. Biomed. Opt. 16(1), 016014 (2011). [CrossRef] [PubMed]
  4. 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]
  5. S. Santos, K. K. Chu, D. Lim, N. Bozinovic, T. N. Ford, C. Hourtoule, A. C. Bartoo, S. K. Singh, and J. Mertz, “Optically sectioned fluorescence endomicroscopy with hybrid-illumination imaging through a flexible fiber bundle,” J. Biomed. Opt. 14(3), 030502 (2009). [CrossRef] [PubMed]
  6. J. W. Goodman, Introduction to Fourier Optics (2nd Ed., McGraw-Hill, 1996), Chap. 6.
  7. C. Ventalon and J. Mertz, “Quasi-confocal fluorescence sectioning with dynamic speckle illumination,” Opt. Lett. 30(24), 3350–3352 (2005). [CrossRef] [PubMed]
  8. D. D. Duncan, S. J. Kirkpatrick, and R. K. Wang, “Statistics of local speckle contrast,” J. Opt. Soc. Am. A 25(1), 9–15 (2008). [CrossRef] [PubMed]
  9. J. Goodman, Speckle phenomena in optics (Roberts & Company, 2007).
  10. J. M. Bélisle, J. P. Correia, P. W. Wiseman, T. E. Kennedy, and S. Costantino, “Patterning protein concentration using laser-assisted adsorption by photobleaching, LAPAP,” Lab Chip 8(12), 2164–2167 (2008). [CrossRef] [PubMed]
  11. J. M. Belisle, D. Kunik, and S. Costantino, “Rapid multicomponent optical protein patterning,” Lab Chip 9(24), 3580–3585 (2009). [CrossRef] [PubMed]
  12. K. Singh, C. Dion, S. Costantino, M. Wajszilber, M. R. Lesk, and T. Ozaki, “Development of a novel instrument to measure the pulsatile movement of ocular tissues,” Exp. Eye Res. 91(1), 63–68 (2010). [CrossRef] [PubMed]
  13. K. Singh, C. Dion, M. R. Lesk, T. Ozaki, and S. Costantino, “Spectral-domain phase microscopy with improved sensitivity using two-dimensional detector arrays,” Rev. Sci. Instrum. 82(2), 023706 (2011). [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

« Previous Article  |  Next Article »

OSA is a member of CrossRef.

CrossCheck Deposited