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Applied Optics

Applied Optics

APPLICATIONS-CENTERED RESEARCH IN OPTICS

  • Editor: James C. Wyant
  • Vol. 45, Iss. 24 — Aug. 20, 2006
  • pp: 6283–6291

Hyperspectral confocal microscope

Michael B. Sinclair, David M. Haaland, Jerilyn A. Timlin, and Howland D. T. Jones  »View Author Affiliations


Applied Optics, Vol. 45, Issue 24, pp. 6283-6291 (2006)
http://dx.doi.org/10.1364/AO.45.006283


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Abstract

We have developed a new, high performance, hyperspectral microscope for biological and other applications. For each voxel within a three-dimensional specimen, the microscope simultaneously records the emission spectrum from 500   nm to 800   nm , with better than 3   nm spectral resolution. The microscope features a fully confocal design to ensure high spatial resolution and high quality optical sectioning. Optical throughput and detection efficiency are maximized through the use of a custom prism spectrometer and a backside thinned electron multiplying charge coupled device (EMCCD) array. A custom readout mode and synchronization scheme enable 512-point spectra to be recorded at a rate of 8300 spectra per second. In addition, the EMCCD readout mode eliminates curvature and keystone artifacts that often plague spectral imaging systems. The architecture of the new microscope is described in detail, and hyperspectral images from several specimens are presented.

OCIS Codes
(180.0180) Microscopy : Microscopy
(180.1790) Microscopy : Confocal microscopy
(180.2520) Microscopy : Fluorescence microscopy

History
Original Manuscript: January 12, 2006
Manuscript Accepted: February 28, 2006

Virtual Issues
Vol. 1, Iss. 9 Virtual Journal for Biomedical Optics

Citation
Michael B. Sinclair, David M. Haaland, Jerilyn A. Timlin, and Howland D. T. Jones, "Hyperspectral confocal microscope," Appl. Opt. 45, 6283-6291 (2006)
http://www.opticsinfobase.org/ao/abstract.cfm?URI=ao-45-24-6283


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References

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  28. The signal-to-noise ratio improvements achieved through simultaneous consideration of all pixels of the image is reduced in the case where a particular component only appears in a small number of pixels, and is weak.

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