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Optics Express

Optics Express

  • Editor: Andrew M. Weiner
  • Vol. 21, Iss. 18 — Sep. 9, 2013
  • pp: 21010–21026

Rapid 3D light-sheet microscopy with a tunable lens

Florian O. Fahrbach, Fabian F. Voigt, Benjamin Schmid, Fritjof Helmchen, and Jan Huisken  »View Author Affiliations


Optics Express, Vol. 21, Issue 18, pp. 21010-21026 (2013)
http://dx.doi.org/10.1364/OE.21.021010


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Abstract

The in-vivo investigation of highly dynamic biological samples, for example the beating zebrafish heart, requires high-speed volume imaging techniques. Light-sheet microscopy is ideal for such samples as it records high-contrast images of entire planes within large samples at once. However, in order to obtain images of different planes, it has been necessary to move the sample relative to the fixed focal plane of the detection objective lens. This mechanical movement limits speed, precision and may be harmful to the sample. We have built a light-sheet microscope that uses remote focusing with an electrically tunable lens (ETL). Without moving specimen or objective we have thereby achieved flexible volume imaging at much higher speeds than previously reported. Our high-speed microscope delivers 3D snapshots of sensitive biological samples. As an example, we imaged 17 planes within a beating zebrafish heart at 510 frames per second, equivalent to 30 volume scans per second. Movements, shape changes and signals across the entire volume can be followed which has been impossible with existing reconstruction techniques.

© 2013 OSA

OCIS Codes
(170.1470) Medical optics and biotechnology : Blood or tissue constituent monitoring
(170.2520) Medical optics and biotechnology : Fluorescence microscopy
(180.6900) Microscopy : Three-dimensional microscopy
(220.1080) Optical design and fabrication : Active or adaptive optics

ToC Category:
Microscopy

History
Original Manuscript: June 24, 2013
Revised Manuscript: August 13, 2013
Manuscript Accepted: August 13, 2013
Published: August 30, 2013

Virtual Issues
Vol. 8, Iss. 10 Virtual Journal for Biomedical Optics

Citation
Florian O. Fahrbach, Fabian F. Voigt, Benjamin Schmid, Fritjof Helmchen, and Jan Huisken, "Rapid 3D light-sheet microscopy with a tunable lens," Opt. Express 21, 21010-21026 (2013)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-21-18-21010


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