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Proceedings Article

Design and analysis of confocal-spectral microscopy using wavelength scanning scheme

[+] Author Affiliations
Dukho Do, Wanhee Chun, Hyeongjun Jeong, Dae-Gab Gweon

KAIST (Korea, Republic of)

Proc. SPIE 7904, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XVIII, 79041L (February 22, 2011); doi:10.1117/12.873667
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From Conference Volume 7904

  • Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XVIII
  • Jose-Angel Conchello; Carol J. Cogswell; Tony Wilson; Thomas G. Brown
  • San Francisco, California, USA | January 22, 2011

abstract

The multi-color, or spectral fluorescence microscopy has ability to detect fluorescence spectral signals which are useful in case of studying interactions and phenomena between biological samples. Recently, commercial devices are combining with confocal microscope so to enhance lateral resolution and to have axial direction discernment. Also Acousto-Optic Tunable Filter(AOTF) is used instead of dichroic mirror to divide excitation and emission signals with mininum light efficiency. In addition, AOTF is used in spectral fluorescence microscopy have many advantages, these are very fast switching speed and high resolution in wavelength selection. However it uses acousto-optic interactions in birefringence material, Tellurium Dioxide(TeO2) , the excitation light interacts with appropriate acoustic signal so that it is diffracted to 1 or -1 order path. But the fluorescence signals from a sample propagate in 0 order path with small different angle according to the polarization state. In this paper, a confocal-spectral microscopy is proposed with the new kind of spectral detector design having wavelength scanning galvano mirror. It makes possible to detect broad wavelength fluorescence signal by single PMT with simply rotating the galvano mirror. Also a new birefringent material, calcite(CaCO3) is used to compensate polarization effect. The proposed spectral confocal microscopy with unique spectrometer body has many advantages in comparison with commercial devices. In terms of detection method, it can be easily applied to other imaging modalities. Hence this system will be adapted in many applications.

© (2011) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Citation

Dukho Do ; Wanhee Chun ; Hyeongjun Jeong and Dae-Gab Gweon
"Design and analysis of confocal-spectral microscopy using wavelength scanning scheme", Proc. SPIE 7904, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XVIII, 79041L (February 22, 2011); doi:10.1117/12.873667; http://dx.doi.org/10.1117/12.873667


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