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Biomedical Optics Express

Biomedical Optics Express

  • Editor: Joseph A. Izatt
  • Vol. 1, Iss. 3 — Oct. 1, 2010
  • pp: 1026–1037

High-speed focal modulation microscopy using acousto-optical modulators

Shau Poh Chong, Chee Howe Wong, Kit Fei Wong, Colin J.R. Sheppard, and Nanguang Chen  »View Author Affiliations

Biomedical Optics Express, Vol. 1, Issue 3, pp. 1026-1037 (2010)

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Focal Modulation Microscopy (FMM) is a single-photon excitation fluorescence microscopy technique which effectively rejects the out-of-focus fluorescence background that arises when imaging deep inside biological tissues. Here, we report on the implementation of FMM in which laser intensity modulation at the focal plane is achieved using acousto-optic modulators (AOM). The modulation speed is greatly enhanced to the MHz range and thus enables real-time image acquisition. The capability of FMM is demonstrated by imaging fluorescence labeled vasculatures in mouse brain as well as self-made tissue phantom.

© 2010 OSA

OCIS Codes
(120.4570) Instrumentation, measurement, and metrology : Optical design of instruments
(170.0170) Medical optics and biotechnology : Medical optics and biotechnology
(180.2520) Microscopy : Fluorescence microscopy
(110.0113) Imaging systems : Imaging through turbid media

ToC Category:

Original Manuscript: May 25, 2010
Revised Manuscript: July 19, 2010
Manuscript Accepted: September 23, 2010
Published: September 30, 2010

Virtual Issues
Advances in Optical Coherence Tomography, Photoacoustic Imaging, and Microscopy (2010) Biomedical Optics Express

Shau Poh Chong, Chee Howe Wong, Kit Fei Wong, Colin J.R. Sheppard, and Nanguang Chen, "High-speed focal modulation microscopy using acousto-optical modulators," Biomed. Opt. Express 1, 1026-1037 (2010)

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  1. J. M. Schmitt, A. Knüttel, and M. Yadlowsky, “Confocal microscopy in turbid media,” J. Opt. Soc. Am. A 11(8), 2226–2235 (1994). [CrossRef] [PubMed]
  2. P. Theer and W. Denk, “On the fundamental imaging-depth limit in two-photon microscopy,” J. Opt. Soc. Am. A 23(12), 3139–3149 (2006). [CrossRef] [PubMed]
  3. J. A. Conchello and J. W. Lichtman, “Optical sectioning microscopy,” Nat. Methods 2(12), 920–931 (2005). [CrossRef] [PubMed]
  4. X. Deng and M. Gu, “Penetration depth of single-, two-, and three-photon fluorescence microscopic imaging through human cortex structures: Monte Carlo simulation,” Appl. Opt. 42(16), 3321–3329 (2003). [CrossRef] [PubMed]
  5. F. Helmchen and W. Denk, “Deep tissue two-photon microscopy,” Nat. Methods 2(12), 932–940 (2005). [CrossRef] [PubMed]
  6. K. Svoboda and R. Yasuda, “Principles of two-photon excitation microscopy and its applications to neuroscience,” Neuron 50(6), 823–839 (2006). [CrossRef] [PubMed]
  7. W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science 248(4951), 73–76 (1990). [CrossRef] [PubMed]
  8. G. H. Patterson and D. W. Piston, “Photobleaching in two-photon excitation microscopy,” Biophys. J. 78(4), 2159–2162 (2000). [CrossRef] [PubMed]
  9. H. F. Zhang, K. Maslov, G. Stoica, and L. V. Wang, “Functional photoacoustic microscopy for high-resolution and noninvasive in vivo imaging,” Nat. Biotechnol. 24(7), 848–851 (2006). [CrossRef] [PubMed]
  10. D. Razansky, M. Distel, C. Vinegoni, R. Ma, N. Perrimon, R. W. Köster, and V. Ntziachristos, “Multispectral opto-acoustic tomography of deep-seated fluorescent proteins in vivo,” Nat. Photonics 3(7), 412–417 (2009). [CrossRef]
  11. C. Ventalon, R. Heintzmann, and J. Mertz, “Dynamic speckle illumination microscopy with wavelet prefiltering,” Opt. Lett. 32(11), 1417–1419 (2007). [CrossRef] [PubMed]
  12. 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]
  13. S. W. Hell and J. Wichmann, “Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy,” Opt. Lett. 19(11), 780–782 (1994). [CrossRef] [PubMed]
  14. N. Chen, C. H. Wong, and C. J. Sheppard, “Focal modulation microscopy,” Opt. Express 16(23), 18764–18769 (2008). [CrossRef] [PubMed]
  15. C. H. Wong, S. P. Chong, C. J. R. Sheppard, and N. Chen, “Simple spatial phase modulator for focal modulation microscopy,” Appl. Opt. 48(17), 3237–3242 (2009). [CrossRef] [PubMed]
  16. S. P. Chong, C. H. Wong, C. J. R. Sheppard, and N. Chen, “Focal modulation microscopy: a theoretical study,” Opt. Lett. 35(11), 1804–1806 (2010). [CrossRef] [PubMed]
  17. Y. Li, Y. Song, L. Zhao, G. Gaidosh, A. M. Laties, and R. Wen, “Direct labeling and visualization of blood vessels with lipophilic carbocyanine dye DiI,” Nat. Protoc. 3(11), 1703–1708 (2008). [CrossRef] [PubMed]
  18. M. Firbank, M. Oda, and D. T. Delpy, “An improved design for a stable and reproducible phantom material for use in near-infrared spectroscopy and imaging,” Phys. Med. Biol. 40(5), 955–961 (1995). [CrossRef] [PubMed]
  19. W. Mo and N. Chen, “Fast time-domain diffuse optical tomography using pseudorandom bit sequences,” Opt. Express 16(18), 13643–13650 (2008). [CrossRef] [PubMed]
  20. S. J. Lunt, C. Gray, C. C. Reyes-Aldasoro, S. J. Matcher, and G. M. Tozer, “Application of intravital microscopy in studies of tumor microcirculation,” J. Biomed. Opt. 15(1), 011113 (2010). [CrossRef] [PubMed]

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