OSA's Digital Library

Optics Letters

Optics Letters


  • Vol. 36, Iss. 17 — Sep. 1, 2011
  • pp: 3503–3505

Three-dimensional in vivo scanning microscopy with inertia-free focus control

Marco Dal Maschio, Angela Michela De Stasi, Fabio Benfenati, and Tommaso Fellin  »View Author Affiliations

Optics Letters, Vol. 36, Issue 17, pp. 3503-3505 (2011)

View Full Text Article

Enhanced HTML    Acrobat PDF (350 KB)

Browse Journals / Lookup Meetings

Browse by Journal and Year


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools



The acquisition of high-resolution images in three dimensions is of utmost importance for the morphological and functional investigation of biological tissues. Here, we present a laser scanning two-photon microscope with remote and motionless control of the focus position. The movement of the excitation spot along the propagation direction is achieved by shaping the laser wavefront with a spatial light modulator. Depending on the optical properties of the objective in use, this approach allows z movements in a range of tens to hundreds of micrometers with small changes of the point spread function. We applied this technique for the three-dimensional (3D) imaging of fluorescent cells in the mouse neocortex in vivo. The presented system bypasses the limitations of microscopes based on moving objectives, enabling high-resolution inertia-free 3D imaging.

© 2011 Optical Society of America

OCIS Codes
(090.1995) Holography : Digital holography
(180.4315) Microscopy : Nonlinear microscopy
(070.6120) Fourier optics and signal processing : Spatial light modulators

ToC Category:
Fourier Optics and Signal Processing

Original Manuscript: July 22, 2011
Manuscript Accepted: August 8, 2011
Published: September 1, 2011

Virtual Issues
Vol. 6, Iss. 10 Virtual Journal for Biomedical Optics

Marco Dal Maschio, Angela Michela De Stasi, Fabio Benfenati, and Tommaso Fellin, "Three-dimensional in vivo scanning microscopy with inertia-free focus control," Opt. Lett. 36, 3503-3505 (2011)

Sort:  Author  |  Year  |  Journal  |  Reset  


  1. A. Diaspro and C. Sheppard, Confocal and Two-Photon Microscopy: Foundations, Applications, and Advances (Wiley-Liss, 2002).
  2. N. Callamaras and I. Parker, Cell Calcium 26, 271 (1999). [CrossRef]
  3. B. Qi, A. P. Himmer, L. M. Gordon, X. D. V. Yang, L. D. Dickensheets, and I. A. Vitkin, Opt. Commun. 232, 123 (2004). [CrossRef]
  4. G. D. Reddy and P. Saggau, J. Biomed. Opt. 10, 064038(2005). [CrossRef]
  5. H. Oku, K. Hashimoto, and M. Ishikawa, Opt. Express 12, 2138 (2004). [CrossRef] [PubMed]
  6. E. J. Botcherby, M. J. Booth, R. Juskaitis, and T. Wilson, Opt. Express 16, 21843 (2008). [CrossRef] [PubMed]
  7. V. R. Daria, C. Stricker, R. Bowman, S. Redman, and H. A. Bachor, Appl. Phys. Lett. 95, 093701 (2009). [CrossRef]
  8. W. Gobel, B. M. Kampa, and F. Helmchen, Nat. Methods 4, 73 (2007). [CrossRef]
  9. A. M. Kerlin, M. L. Andermann, V. K. Berezovskii, and R. C. Reid, Neuron 67, 858 (2010). [CrossRef] [PubMed]
  10. A. Cheng, J. T. Goncalves, P. Golshani, K. Arisaka, and C. Portera-Cailliau, Nat. Methods 8, 139 (2011). [CrossRef] [PubMed]
  11. M. Dal Maschio, F. Difato, R. Beltramo, A. Blau, F. Benfenati, and T. Fellin, Opt. Express 18, 18720 (2010). [CrossRef] [PubMed]
  12. V. Nikolenko, B. O. Watson, R. Araya, A. Woodruff, D. S. Peterka, and R. Yuste, Front. Neural Circuits 2, 5 (2008). [CrossRef]
  13. J. E. Curtis, B. A. Koss, and D. G. Grier, Opt. Commun. 207, 169 (2002). [CrossRef]
  14. V. Emiliani, D. Cojoc, E. Ferrari, V. Garbin, C. Durieux, M. Coppey-Moisan, and F. E. Di, Opt. Express 13, 1395 (2005). [CrossRef] [PubMed]
  15. O. Garaschuk, R. I. Milos, and A. Konnerth, Nat. Protoc. 1, 380 (2006). [CrossRef]
  16. R. G. Duemani, K. Kelleher, R. Fink, and P. Saggau, Nat. Neurosci. 11, 713 (2008). [CrossRef]
  17. B. F. Grewe, D. Langer, H. Kasper, B. M. Kampa, and F. Helmchen, Nat. Methods 7, 399 (2010). [CrossRef] [PubMed]
  18. B. F. Grewe, F. F. Voigt, H. M. van’t Hoff, and F. Helmchen, Biomed. Opt. Express 2, 2035 (2011). [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.


Fig. 1 Fig. 2 Fig. 3

« Previous Article  |  Next Article »

OSA is a member of CrossRef.

CrossCheck Deposited