Rapid adaptive optical recovery of optimal resolution over large volumes
Abstract
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.
Additional Information
© 2014 Macmillan Publishers Limited. Received 9 December 2013 ; Accepted 6 March 2014 ; Published online 13 April 2014. 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 CIPS M) 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. Author Contributions: E.B. supervised the project; K.W. and E.B. conceived the idea; D.E.M., K.W. and E.B. developed the instrument control program; K.W. built the instrument and performed the experiments; A.S., P.E., T.M., M.E.B. and J.M. supplied zebrafish lines and guidance on live zebrafish imaging; K.W. and E.B. analyzed the data; E.B. wrote the paper with input from all co-authors.Attached Files
Accepted Version - nihms576781.pdf
Supplemental Material - nmeth.2925-S1.pdf
Supplemental Material - nmeth.2925-sv1.mov
Supplemental Material - nmeth.2925-sv2.mov
Supplemental Material - nmeth.2925-sv3.mov
Supplemental Material - nmeth.2925-sv4.mov
Supplemental Material - nmeth.2925-sv5.mov
Supplemental Material - nmeth.2925-sv6.mov
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Additional details
- PMCID
- PMC4069208
- Eprint ID
- 44076
- Resolver ID
- CaltechAUTHORS:20140303-060128636
- R21 MH083614
- National Institute of Mental Health (NIMH)
- R43 HD047089
- National Institute of Child Health and Human Development (NICHD)
- DE16459
- NIH
- EXC114 CIPS M
- Center for Integrated Protein Sciences
- EXC1010 SyNergy
- Munich Cluster for Systems Neurology
- Deutsche Forschungsgemeinschaft (DFG)
- Howard Hughes Medical Institute (HHMI)
- Created
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2014-04-14Created from EPrint's datestamp field
- Updated
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2021-11-10Created from EPrint's last_modified field