OSA's Digital Library

Biomedical Optics Express

Biomedical Optics Express

  • Editor: Joseph A. Izatt
  • Vol. 1, Iss. 1 — Aug. 2, 2010
  • pp: 236–245

Practical implementation of log-scale active illumination microscopy

Kengyeh K. Chu, Daryl Lim, and Jerome Mertz  »View Author Affiliations

Biomedical Optics Express, Vol. 1, Issue 1, pp. 236-245 (2010)

View Full Text Article

Enhanced HTML    Acrobat PDF (1486 KB)

Browse Journals / Lookup Meetings

Browse by Journal and Year


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools



Active illumination microscopy (AIM) is a method of redistributing dynamic range in a scanning microscope using real-time feedback to control illumination power on a sub-pixel time scale. We describe and demonstrate a fully integrated instrument that performs both feedback and image reconstruction. The image is reconstructed on a logarithmic scale to accommodate the dynamic range benefits of AIM in a single output channel. A theoretical and computational analysis of the influence of noise on active illumination feedback is presented, along with imaging examples illustrating the benefits of AIM. While AIM is applicable to any type of scanning microscope, we apply it here specifically to two-photon microscopy.

© 2010 Optical Society of America

OCIS Codes
(110.0180) Imaging systems : Microscopy
(180.2520) Microscopy : Fluorescence microscopy
(180.5810) Microscopy : Scanning microscopy
(190.4180) Nonlinear optics : Multiphoton processes

ToC Category:

Original Manuscript: May 28, 2010
Revised Manuscript: July 7, 2010
Manuscript Accepted: July 7, 2010
Published: July 16, 2010

Kengyeh K. Chu, Daryl Lim, and Jerome Mertz, "Practical implementation of log-scale active illumination microscopy," Biomed. Opt. Express 1, 236-245 (2010)

Sort:  Author  |  Year  |  Journal  |  Reset  


  1. J. B. Pawley, Handbook of biological confocal microscopy (Springer, New York, 2006).
  2. W. Denk, J. H. Strickler, and W. W. Webb, "Two-photon laser scanning fluuorescence microscopy," Science 248, 73-76 (1990).
  3. R. A. Hoebe, C. H. Van Oven, T. W. J. GadellaJr., P. B. Dhonukshe, C. J. F. Van Noorden, and E. M. M. Manders, "Controlled light-exposure microscopy reduces photobleaching and phototoxicity in fluorescence livecell imaging," Nat. Biotechnol. 25,249-253 (2007).
  4. K. K. Chu, D. Lim, and J. Mertz, "Enhanced weak-signal sensitivity in two-photon microscopy by adaptive illumination," Opt. Lett. 32, 2846-2848 (2007).
  5. G. H. Patterson and D. W. Piston, "Photobleaching in two-photon excitation microscopy," Biophys. J. 78, 2159-2162 (2000).
  6. M. Steinbauer, A. G. Harris, C. Abels, and K. Messmer, "Charactierization and prevention of phototoxic effects intravital fluorescence microscopy in the hamster dorsal skinfold model," Langenbecks Arch. Surg. 385, 290-298 (2000).
  7. I. Navarro-Quiroga, R. Chittajalu, V. Gallo, and T. F. Haydar, "Long-term, selective gene expression in developing and adult hippocampal pyramidal neurons using focal in utero electroporation," J. Neurosci. 27, 5007-5011 (2007).

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.

Supplementary Material

» Media 1: AVI (2344 KB)     

« Previous Article  |  Next Article »

OSA is a member of CrossRef.

CrossCheck Deposited