Adaptive movement compensation for in vivo imaging of fast cellular dynamics within a moving tissue.

TitleAdaptive movement compensation for in vivo imaging of fast cellular dynamics within a moving tissue.
Publication TypeJournal Article
Year of Publication2011
AuthorsLaffray S, Pagès S, Dufour H, De Koninck P, De Koninck Y, Côté D
JournalPLoS One
Volume6
Issue5
Paginatione19928
Date Published2011
ISSN1932-6203
KeywordsAnimals, Calcium, Diagnostic Imaging, Male, Rats, Rats, Wistar, Spinal Cord
Abstract

In vivo non-linear optical microscopy has been essential to advance our knowledge of how intact biological systems work. It has been particularly enabling to decipher fast spatiotemporal cellular dynamics in neural networks. The power of the technique stems from its optical sectioning capability that in turn also limits its application to essentially immobile tissue. Only tissue not affected by movement or in which movement can be physically constrained can be imaged fast enough to conduct functional studies at high temporal resolution. Here, we show dynamic two-photon Ca(2+) imaging in the spinal cord of a living rat at millisecond time scale, free of motion artifacts using an optical stabilization system. We describe a fast, non-contact adaptive movement compensation approach, applicable to rough and weakly reflective surfaces, allowing real-time functional imaging from intrinsically moving tissue in live animals. The strategy involves enslaving the position of the microscope objective to that of the tissue surface in real-time through optical monitoring and a closed feedback loop. The performance of the system allows for efficient image locking even in conditions of random or irregular movements.

DOI10.1371/journal.pone.0019928
Alternate JournalPLoS ONE
PubMed ID21629702
PubMed Central IDPMC3101223
Grant List / / Canadian Institutes of Health Research / Canada

Funding

Our research endeavors are made possible by the following agencies:

Canadian Institutes of Health Research - Instituts de recherche en santé du Canada Fonds de recherche du Québec – Nature et technologies (FRQNT)Fonds de la recherche en santé du Québec   Natural Sciences and Engineering Research Council of Canada (NSERC) - Conseil de recherche en sciences naturelles et en génie du Canada (CRSNG)innovation.caHuman Frontier Science ProgramCanada First Research Excellence FundSentinelle Nord