OSA's Digital Library

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)

View Full Text Article

Enhanced HTML    Acrobat PDF (19160 KB)

Browse Journals / Lookup Meetings

Browse by Journal and Year


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools



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:

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

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)

Sort:  Author  |  Year  |  Journal  |  Reset  


  1. S.  Ram, P.  Prabhat, J.  Chao, E. S.  Ward, R. J.  Ober, “High Accuracy 3D Quantum Dot Tracking with Multifocal Plane Microscopy for the Study of Fast Intracellular Dynamics in Live Cells,” Biophys. J. 95, 6025–6043 (2008). [CrossRef] [PubMed]
  2. S.  Abrahamsson, J.  Chen, B.  Hajj, S.  Stallinga, A. Y.  Katsov, J.  Wisniewski, G.  Mizuguchi, P.  Soule, F.  Mueller, C.  Dugast Darzacq, X.  Darzacq, C.  Wu, C. I.  Bargmann, D. A.  Agard, M.  Dahan, M. G. L.  Gustafsson, “Fast multicolor 3D imaging using aberration-corrected multifocus microscopy,” Nat. Methods 10, 60–63 (2012). [CrossRef] [PubMed]
  3. P. A.  Dalgarno, H. I. C.  Dalgarno, A.  Putoud, R.  Lambert, L.  Paterson, D. C.  Logan, D. P.  Towers, R. J.  Warburton, A. H.  Greenaway, “Multiplane imaging and three dimensional nanoscale particle tracking in biological microscopy,” Opt. Express 18, 877–884 (2010). [CrossRef] [PubMed]
  4. M.  Levoy, Z.  Zhang, I.  McDowall, “Recording and controlling the 4D light field in a microscope using microlens arrays,” J. Mic. 235, 144–162 (2009). [CrossRef]
  5. J.  Huisken, J.  Swoger, F.  Del Bene, J.  Wittbrodt, E. H. K.  Stelzer, “Optical sectioning deep inside live embryos by selective plane illumination microscopy,” Science 305, 1007–1009 (2004). [CrossRef] [PubMed]
  6. J.  Huisken, D. Y. R.  Stainier, “Selective plane illumination microscopy techniques in developmental biology,” Development 136, 1963–1975 (2009). [CrossRef] [PubMed]
  7. A.  Kaufmann, M.  Mickoleit, M.  Weber, J.  Huisken, “Multilayer mounting enables long-term imaging of zebrafish development in a light sheet microscope,” Development 139, 3242–3247 (2012). [CrossRef] [PubMed]
  8. T.  Panier, S.  Romano, R.  Olive, T.  Pietri, G.  Sumbre, R.  Candelier, G.  Debrégeas, “Fast functional imaging of multiple brain regions in intact zebrafish larvae using Selective Plane Illumination Microscopy,” Front. Neural Circuits 7, 65 (2013). [CrossRef] [PubMed]
  9. T. F.  Holekamp, D.  Turaga, T. E.  Holy, “Fast Three-Dimensional Fluorescence Imaging of Activity in Neural Populations by Objective-Coupled Planar Illumination Microscopy,” Neuron 57, 661–672 (2008). [CrossRef] [PubMed]
  10. S.  Kumar, D.  Wilding, M. B.  Sikkel, A. R.  Lyon, K. T.  MacLeod, C.  Dunsby, “High-speed 2D and 3D fluorescence microscopy of cardiac myocytes,” Opt. Express 19, 13839–13847 (2011). [CrossRef] [PubMed]
  11. C.  Dunsby, “Optically sectioned imaging by oblique plane microscopy,” Opt. Express 16, 20306–20316 (2009). [CrossRef]
  12. E. J.  Botcherby, R.  Juškaitis, M. J.  Booth, T.  Wilson, “An optical technique for remote focusing in microscopy,” Opt. Commun. 281, 880–887 (2008). [CrossRef]
  13. E. J.  Botcherby, C. W.  Smith, M. M.  Kohl, D.  Débarre, M. J.  Booth, R.  Juškaitis, O.  Paulsen, T.  Wilson, “Aberration-free three-dimensional multiphoton imaging of neuronal activity at kHz rates,” P. Natl Acad. Sci. USA 109, 2919–2924 (2012). [CrossRef]
  14. J.  Huisken, D. Y. R.  Stainier, “Even fluorescence excitation by multidirectional selective plane illumination microscopy (mSPIM),” Opt. Lett. 32, 2608–2610 (2007). [CrossRef] [PubMed]
  15. B. F.  Grewe, F. F.  Voigt, M.  van’t Hoff, F.  Helmchen, “Fast two-layer two-photon imaging of neuronal cell populations using an electrically tunable lens,” Biomed. Opt. Exp. 2, 2035–2046 (2011). [CrossRef]
  16. M.  Liebling, A. S.  Forouhar, R.  Wolleschensky, B.  Zimmermann, R.  Ankerhold, S. E.  Fraser, M.  Gharib, M. E.  Dickinson, “Rapid three-dimensional imaging and analysis of the beating embryonic heart reveals functional changes during development,” Dev. Dynam. 235, 2940–2948 (2006). [CrossRef]
  17. C.  Skupsch, C.  Brücker, “Multiple-plane particle image velocimetry using a light-field camera,” Opt. Express 21, 1726–1740 (2013). [CrossRef] [PubMed]

Cited By

Alert me when this paper is cited

OSA is able to provide readers links to articles that cite this paper by participating in CrossRef's Cited-By Linking service. CrossRef includes content from more than 3000 publishers and societies. In addition to listing OSA journal articles that cite this paper, citing articles from other participating publishers will also be listed.

Supplementary Material

» Media 1: MOV (11580 KB)     
» Media 2: MOV (894 KB)     
» Media 3: MOV (2512 KB)     
» Media 4: MOV (8427 KB)     

« Previous Article  |  Next Article »

OSA is a member of CrossRef.

CrossCheck Deposited