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Original Research ARTICLE

SLM microscopy: scanless two-photon imaging and photostimulation with spatial light modulators

Volodymyr Nikolenko, Brendon O. Watson, Roberto Araya, Alan Woodruff, Darcy S. Peterka and Rafael Yuste^*

Department of Biological Sciences, Howard Hughes Medical Institute, Columbia University, New York, NY, USA

Laser microscopy has generally poor temporal resolution, caused by the serial scanning of each pixel. This is a significant problem for imaging or optically manipulating neural circuits, since neuronal activity is fast. To help surmount this limitation, we have developed a “scanless” microscope that does not contain mechanically moving parts. This microscope uses a diffractive spatial light modulator (SLM) to shape an incoming two-photon laser beam into any arbitrary light pattern. This allows the simultaneous imaging or photostimulation of different regions of a sample with three-dimensional precision. To demonstrate the usefulness of this microscope, we perform two-photon uncaging of glutamate to activate dendritic spines and cortical neurons in brain slices. We also use it to carry out fast (60 Hz) two-photon calcium imaging of action potentials in neuronal populations. Thus, SLM microscopy appears to be a powerful tool for imaging and optically manipulating neurons and neuronal circuits. Moreover, the use of SLMs expands the flexibility of laser microscopy, as it can substitute traditional simple fixed lenses with any calculated lens function.

Keywords:

spines, DOE, MNI-glutamate cortex

Citation:

Nikolenko V, Watson BO, Araya R, Woodruff A, Peterka DS and Yuste R (2008). SLM microscopy: scanless two-photon imaging and photostimulation with spatial light modulators. Front. Neural Circuits 2 :5. doi: 10.3389/neuro.04.005.2008

Received:

25 September 2008;

Paper pending published:

11 October 2008;

Accepted:

19 November 2008;

Published online:

19 December 2008.

Edited by:

Rachel O. Wong, University of Washington, USA

Reviewed by:

Karl Deisseroth, Stanford University, USA
Tim Holy, Washington University School of Medicine, USA

© 2008 Nikolenko, Watson, Araya, Woodruff, Peterka 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