Ultrafast random-access scanning in two-photon microscopy using acousto-optic deflectors

Autor: Jean-François Léger, Oleg Krichevsky, Claire Wyart, Stéphane Dieudonné, D. Chatenay, Yves Kremer, R. Salomé, Laurent Bourdieu
Přispěvatelé: Laboratoire de Dynamique des Fluides Complexes (LDFC), Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Neurobiologie (UMR 8544) (NEURO), École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Physique Théorique de l'ENS (LPTENS), Université Pierre et Marie Curie - Paris 6 (UPMC)-Fédération de recherche du Département de physique de l'Ecole Normale Supérieure - ENS Paris (FRDPENS), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS Paris)-Institut National de la Santé et de la Recherche Médicale (INSERM), Fédération de recherche du Département de physique de l'Ecole Normale Supérieure - ENS Paris (FRDPENS), Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS Paris)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS Paris)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), École normale supérieure - Paris (ENS-PSL), Laboratoire de Physique Théorique de l'ENS [École Normale Supérieure] (LPTENS), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)
Jazyk: angličtina
Rok vydání: 2006
Předmět:
MESH: Hippocampus
[SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology
MESH: Neurons
Action Potentials
Field of view
MESH: Microscopy
Fluorescence
MESH: Signal Processing
Computer-Assisted
01 natural sciences
Hippocampus
Two-photon excitation microscopy
MESH: Microscopy
Confocal
MESH: Animals
MESH: Models
Theoretical
Image resolution
Cells
Cultured
MESH: Action Potentials
Physics
Neurons
0303 health sciences
Millisecond
Microscopy
Confocal
General Neuroscience
Pyramidal Cells
Resolution (electron density)
Signal Processing
Computer-Assisted
Femtosecond
MESH: Lasers
Algorithms
MESH: Cells
Cultured
Diagnostic Imaging
MESH: Rats
MESH: Algorithms
MESH: Calcium Signaling
010309 optics
03 medical and health sciences
Optics
0103 physical sciences
Animals
Calcium Signaling
030304 developmental biology
business.industry
MESH: Diagnostic Imaging
Lasers
MESH: Pyramidal Cells
Models
Theoretical
Rats
Microscopy
Fluorescence
Temporal resolution
business
Ultrashort pulse
Zdroj: Journal of Neuroscience Methods
Journal of Neuroscience Methods, Elsevier, 2006, 154 (1-2), pp.161-74. ⟨10.1016/j.jneumeth.2005.12.010⟩
Journal of Neuroscience Methods, 2006, 154 (1-2), pp.161-74. ⟨10.1016/j.jneumeth.2005.12.010⟩
ISSN: 0165-0270
Popis: Two-photon scanning microscopy (TPSM) is a powerful tool for imaging deep inside living tissues with sub-cellular resolution. The temporal resolution of TPSM is however strongly limited by the galvanometric mirrors used to steer the laser beam. Fast physiological events can therefore only be followed by scanning repeatedly a single line within the field of view. Because acousto-optic deflectors (AODs) are non-mechanical devices, they allow access at any point within the field of view on a microsecond time scale and are therefore excellent candidates to improve the temporal resolution of TPSM. However, the use of AOD-based scanners with femtosecond pulses raises several technical difficulties. In this paper, we describe an all-digital TPSM setup based on two crossed AODs. It includes in particular an acousto-optic modulator (AOM) placed at 45 degrees with respect to the AODs to pre-compensate for the large spatial distortions of femtosecond pulses occurring in the AODs, in order to optimize the spatial resolution and the fluorescence excitation. Our setup allows recording from freely selectable point-of-interest at high speed (1kHz). By maximizing the time spent on points of interest, random-access TPSM (RA-TPSM) constitutes a promising method for multiunit recordings with millisecond resolution in biological tissues.
Databáze: OpenAIRE
Externí odkaz: