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Original Research ARTICLE

Two-photon imaging with diffractive optical elements

Department of Biological Sciences, Howard Hughes Medical Institute, Columbia University, New York, NY, USA
Two-photon imaging has become a useful tool for optical monitoring of neural circuits, but it requires high laser power and serial scanning of each pixel in a sample. This results in slow imaging rates, limiting the measurements of fast signals such as neuronal activity. To improve the speed and signal-to-noise ratio of two-photon imaging, we introduce a simple modification of a two-photon microscope, using a diffractive optical element (DOE) which splits the laser beam into several beamlets that can simultaneously scan the sample. We demonstrate the advantages of DOE scanning by enhancing the speed and sensitivity of two-photon calcium imaging of action potentials in neurons from neocortical brain slices. DOE scanning can easily improve the detection of time-varying signals in two-photon and other non-linear microscopic techniques.
Keywords:
beam-splitter, video-rate, calcium, scanning, uncaging, photostimulation
Citation:
Watson BO, Nikolenko V and Yuste R (2009). Two-photon imaging with diffractive optical elements. Front. Neural Circuits 3:6. doi: 10.3389/neuro.04.006.2009
Received:
10 February 2009;
 Paper pending published:
09 March 2009;
Accepted:
15 June 2009;
 Published online:
06 July 2009.

Edited by:

Gero Miesenboeck, Oxford University, UK

Reviewed by:

Fritjof Helmchen, University of Zurich, Switzerland
Alan Fine, Dalhousie University, Canada
Copyright:
© 2009 Watson, Nikolenko and Yuste. This is an open-access article subject to an exclusive license agreement between the authors and the Frontiers Research Foundation, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are credited.
*Correspondence:
Rafael Yuste, Department of Biological Sciences, Columbia University, 1212 Amsterdam Avenue, Box 2435, New York, NY 10027, USA. e-mail: rafaelyuste@columbia.edu

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