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Biomedical Optics Express

Biomedical Optics Express

  • Editor: Joseph A. Izatt
  • Vol. 2, Iss. 7 — Jul. 1, 2011
  • pp: 2035–2046

Fast two-layer two-photon imaging of neuronal cell populations using an electrically tunable lens

Benjamin F. Grewe, Fabian F. Voigt, Marcel van ’t Hoff, and Fritjof Helmchen  »View Author Affiliations


Biomedical Optics Express, Vol. 2, Issue 7, pp. 2035-2046 (2011)
http://dx.doi.org/10.1364/BOE.2.002035


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Abstract

Functional two-photon Ca2+-imaging is a versatile tool to study the dynamics of neuronal populations in brain slices and living animals. However, population imaging is typically restricted to a single two-dimensional image plane. By introducing an electrically tunable lens into the excitation path of a two-photon microscope we were able to realize fast axial focus shifts within 15 ms. The maximum axial scan range was 0.7 mm employing a 40x NA0.8 water immersion objective, plenty for typically required ranges of 0.2–0.3 mm. By combining the axial scanning method with 2D acousto-optic frame scanning and random-access scanning, we measured neuronal population activity of about 40 neurons across two imaging planes separated by 40 μm and achieved scan rates up to 20–30 Hz. The method presented is easily applicable and allows upgrading of existing two-photon microscopes for fast 3D scanning.

© 2011 OSA

OCIS Codes
(170.0180) Medical optics and biotechnology : Microscopy
(170.2520) Medical optics and biotechnology : Fluorescence microscopy
(180.6900) Microscopy : Three-dimensional microscopy
(180.4315) Microscopy : Nonlinear microscopy

ToC Category:
Microscopy

History
Original Manuscript: March 28, 2011
Revised Manuscript: June 10, 2011
Manuscript Accepted: June 10, 2011
Published: June 23, 2011

Citation
Benjamin F. Grewe, Fabian F. Voigt, Marcel van ’t Hoff, and Fritjof Helmchen, "Fast two-layer two-photon imaging of neuronal cell populations using an electrically tunable lens," Biomed. Opt. Express 2, 2035-2046 (2011)
http://www.opticsinfobase.org/boe/abstract.cfm?URI=boe-2-7-2035


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