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Optics Express

Optics Express

  • Editor: Andrew M. Weiner
  • Vol. 21, Iss. 20 — Oct. 7, 2013
  • pp: 23611–23618

Development of a high speed laser scanning confocal microscope with an acquisition rate up to 200 frames per second

S. Choi, P. Kim, R. Boutilier, M. Y. Kim, Y. J. Lee, and H. Lee  »View Author Affiliations

Optics Express, Vol. 21, Issue 20, pp. 23611-23618 (2013)

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There has been an increasing interest for observing fast biological phenomena such as cell movements in circulations and action potentials. The laser scanning confocal microscopy offers a good spatial resolution and optical sectioning ability to observe various in vivo animal models. We developed a high speed laser scanning confocal microscope capable of acquiring 512 by 512 pixel images at 200 fps (frames per second). We have incorporated a fast rotating polygonal scanning mirror with 128 facets for the X-axis scanner. In order to increase the throughput of the Y-axis scanner, we applied a bi-directional scanning method for vertical scanning. This made it possible to scan along the Y-axis two times during each scanner motion cycle. For the image acquisition, we used a custom photomultiplier tube amplifier with a broad frequency band. In addition, custom imaging software was written for the new microscope. In order to verify the acquisition speed of the developed confocal microscope, a resolution target moving at a series of constant speeds and a sedated mouse with slight movements due to heartbeats were observed. By comparing successive frames, the frame acquisition speeds were calculated.

© 2013 OSA

OCIS Codes
(120.4570) Instrumentation, measurement, and metrology : Optical design of instruments
(170.1790) Medical optics and biotechnology : Confocal microscopy
(170.5810) Medical optics and biotechnology : Scanning microscopy

ToC Category:

Original Manuscript: August 13, 2013
Revised Manuscript: September 9, 2013
Manuscript Accepted: September 10, 2013
Published: September 26, 2013

S. Choi, P. Kim, R. Boutilier, M. Y. Kim, Y. J. Lee, and H. Lee, "Development of a high speed laser scanning confocal microscope with an acquisition rate up to 200 frames per second," Opt. Express 21, 23611-23618 (2013)

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