Using a descanned, laser-induced guide star and direct wavefront sensing, we demonstrate adaptive correction of complex optical aberrations at high numerical aperture (NA) and a 14-ms update rate. This correction permits us to compensate for the rapid spatial variation in aberration often encountered in biological specimens and to recover diffraction-limited imaging over large volumes (>240 mm per side). We applied this to image fine neuronal processes and subcellular dynamics within the zebrafish brain.
This is a preview of subscription content, access via your institution
Open Access articles citing this article.
Nature Methods Open Access 28 September 2023
Communications Biology Open Access 31 January 2023
Biomaterials Research Open Access 22 October 2022
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Rent or buy this article
Prices vary by article type
Prices may be subject to local taxes which are calculated during checkout
Hardy, J.W. Adaptive Optics for Astronomical Telescopes (Oxford Univ. Press, 1998).
Booth, M.J. Phil. Trans. R. Soc. A 365, 2829–2843 (2007).
Kubby, J.A. Adaptive Optics for Biological Imaging (CRC Press, 2013).
Schwertner, M., Booth, M.J. & Wilson, T. Opt. Express. 12, 6540–6552 (2004).
Aviles-Espinosa, R. et al. Biomed. Opt. Express 2, 3135–3149 (2011).
Hofer, H., Artal, P., Singer, B., Aragón, J.L. & Williams, D.R. J. Opt. Soc. Am. A Opt. Image Sci. Vis. 18, 497–506 (2001).
Tao, X. et al. Opt. Lett. 36, 1062–1064 (2011).
Tao, X. et al. Opt. Lett. 36, 3389–3391 (2011).
Débarre, D. et al. Opt. Lett. 34, 2495–2497 (2009).
Ji, N., Milkie, D.E. & Betzig, E. Nat. Methods 7, 141–147 (2010).
Cui, M. Opt. Lett. 36, 870–872 (2011).
Milkie, D.E., Betzig, E. & Ji, N. Opt. Lett. 36, 4206–4208 (2011).
Keller, P.J., Schmidt, A.D., Wittbrodt, J. & Stelzer, E.H.K. Science 322, 1065–1069 (2008).
Kaufmann, A., Mickoleit, M., Weber, M. & Husiken, J. Development 139, 3242–3247 (2012).
Tomer, R., Khairy, K. & Keller, P.J. Curr. Opin. Genet. Dev. 21, 558–565 (2011).
Weber, M. & Huisken, J. Curr. Opin. Genet. Dev. 21, 566–572 (2011).
Planchon, T.A. et al. Nat. Methods 8, 417–423 (2011).
Gao, L. et al. Cell 151, 1370–1385 (2012).
Ahrens, M.B. et al. Nature 485, 471–477 (2012).
Ahrens, M.B. et al. Nat. Methods 10, 413–420 (2013).
Gerchberg, R.W. & Saxton, W.O. Optik (Stuttg.) 35, 237–246 (1972).
Campbell, H.I., Zhang, S., Greenaway, A.H. & Restaino, S. Opt. Lett. 29, 2707–2709 (2004).
Kimmel, C.B., Ballard, W.W., Kimmel, S.R., Ullmann, B. & Schilling, T.F. Dev. Dyn. 203, 253–310 (1995).
Westerfield, M. The Zebrafish Book: A Guide for the Laboratory Use of Zebrafish (Danio rerio) 4th edn. (University of Oregon Press, 2000).
Xie, X. et al. BMC Biol. 10, 93 (2012).
Cooper, M.S. et al. Dev. Dyn. 232, 359–368 (2005).
Wada, N. et al. Development 132, 3977–3988 (2005).
Carney, T.J. et al. Development 133, 4619–4630 (2006).
Blader, P., Plessy, C. & Strahle, U. Mech. Dev. 120, 211–218 (2003).
Szobota, S. et al. Neuron 54, 535–545 (2007).
Plucińska, G. et al. J. Neurosci. 32, 16203–16212 (2012).
Godinho, L. in Imaging in Developmental Biology (eds. Sharpe, J. & Wong, R.O.) Ch. 4, 49–69 (Cold Spring Harbor Laboratory Press, 2011).
We thank our colleagues N. Ji for many fruitful technical discussions and suggestion of the zebrafish system; P. Keller for the HRAS transgenic line; C. Yang, S. Narayan, M.B. Ahrens, M. Koyama, B. Lemon, K. McDole and P. Keller for further guidance on zebrafish biology; J. Cox, M. Rose, A. Luck and J. Barber for zebrafish maintenance and breeding; and R. Kloss, B. Biddle and B. Bowers for machining services. We are grateful to R. Köster (Technical University of Braunschweig) for providing the KalTA4 transactivator and X. Xie (Georgia Regents University) for assistance in generating corresponding transgenic Enhancer Trap lines. We also thank R. Kelsh (University of Bath) for the Sox10:eGFP line and U. Strahle (Karlsruhe Institute of Technology) for the Ngn:nRFP line. J.S.M. is supported by US National Institutes of Health (NIH) grants R21 MH083614 (NIMH) and R43 HD047089 (NICHD). M.E.B. is supported by NIH grant DE16459. T.M. acknowledges the financial support of the Center for Integrated Protein Sciences (EXC114 CIPSM) and of the Munich Cluster for Systems Neurology (EXC1010 SyNergy). P.E. was supported by DFG Research Training Group 1373. A.S. and P.E. acknowledge support from the Howard Hughes Medical Institute Janelia Farm visiting scientist program.
The authors declare no competing financial interests.
Supplementary Figures 1–9 and Supplementary Tables 1 and 2 (PDF 3774 kb)
Two photon 3D image of a membrane-labeled subset of neurons in the brain of a living zebrafish embryo, 72 hours post fertilization. This 240 x 240 x 270 μm3 imaging volume consists of 19,584 corrective subvolumes, each 30 x 30 x 1.05 μm3 in extent. Zooming deep in the mid-brain, the individual neuronal processes, unresolved without AO, become distinct after correction. (MOV 49041 kb)
Two-photon time lapse imaging of the neurite-guided oligodendrocyte migration deep in the zebrafish hindbrain with adaptive optical correction. All frames are MIPs over a 170 x 90 x 60 μm volume taken at 4 minute intervals with AO and deconvolution in the two-photon mode, starting ∼72 hours post-fertilization. (MOV 3951 kb)
Comparative two-photon imaging of a living zebrafish brain, ∼72 hours post-fertilization, with a ubiquitously expressed cell membrane marker at a depth of 150 μm. Left panel: no AO; middle panel: with AO; right panel: local wavefront error. (MOV 27099 kb)
Two–color, 3D confocal images of oligodendrocytes and neuronal nuclei over a 40 x 40 x 200 μm3 volume extending from the optic tectum through the midbrain, in a zebrafish 72 hours post fertilization. AO correction was performed using a de-scanned two-photon guide star in each of 40 x 40 x 9 μm3 corrective sub-volumes before confocal imaging. (MOV 24581 kb)
Two–color, 3D confocal images of mitochondria (magenta) and the plasma membrane (green) of a cell ∼150 μm deep in the hindbrain of a zebrafish, 4 days post fertilization. The imaging volume is 25 x 25 x 15 μm3. (MOV 10896 kb)
Time lapse imaging of axonal trafficking of mitochondria 150 μm deep in the zebrafish brain, 4 days post fertilization, over a 20 x 40 x 18 μm3 imaging volume at 2 minute intervals. (MOV 8122 kb)
About this article
Cite this article
Wang, K., Milkie, D., Saxena, A. et al. Rapid adaptive optical recovery of optimal resolution over large volumes. Nat Methods 11, 625–628 (2014). https://doi.org/10.1038/nmeth.2925
This article is cited by
Communications Biology (2023)
Nature Methods (2023)
Biomaterials Research (2022)
Nature Reviews Nephrology (2022)
Nature Biotechnology (2022)