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Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics


  • Editor: Gregory W. Faris
  • Vol. 2, Iss. 9 — Sep. 26, 2007

Multifocal multiphoton microscopy (MMM) at a frame rate beyond 600 Hz

Karsten Bahlmann, Peter T. C. So, Michael Kirber, Robert Reich, Bernard Kosicki, William McGonagle, and Karl Bellve  »View Author Affiliations

Optics Express, Vol. 15, Issue 17, pp. 10991-10998 (2007)

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We introduce a multiphoton microscope for high-speed three-dimensional (3D) fluorescence imaging. The system combines parallel illumination by a multifocal multiphoton microscope (MMM) with parallel detection via a segmented high-sensitivity charge-couple device (CCD) camera. The instrument consists of a Ti-sapphire laser illuminating a microlens array that projects 36 foci onto the focal plane. The foci are scanned using a resonance scanner and imaged with a custom-made CCD camera. The MMM increases the imaging speed by parallelizing the illumination; the CCD camera can operate at a frame rate of 1428 Hz while maintaining a low read noise of 11 electrons per pixel by dividing its chip into 16 independent segments for parallelized readout. We image fluorescent specimens at a frame rate of 640 Hz. The calcium wave of fluo3 labeled cardiac myocytes is measured by imaging the spontaneous contraction of the cells in a 0.625 second sequence movie, consisting of 400 single images.

© 2007 Optical Society of America

OCIS Codes
(180.0180) Microscopy : Microscopy
(180.2520) Microscopy : Fluorescence microscopy
(180.5810) Microscopy : Scanning microscopy
(180.6900) Microscopy : Three-dimensional microscopy
(190.0190) Nonlinear optics : Nonlinear optics
(190.4180) Nonlinear optics : Multiphoton processes

ToC Category:

Original Manuscript: July 16, 2007
Revised Manuscript: August 14, 2007
Manuscript Accepted: August 14, 2007
Published: August 16, 2007

Virtual Issues
Vol. 2, Iss. 9 Virtual Journal for Biomedical Optics

Karsten Bahlmann, Peter T. So, Michael Kirber, Robert Reich, Bernard Kosicki, William McGonagle, and Karl Bellve, "Multifocal multiphoton microscopy (MMM) at a frame rate beyond 600 Hz," Opt. Express 15, 10991-10998 (2007)

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