TY - JOUR
T1 - Ultrafast random-access scanning in two-photon microscopy using acousto-optic deflectors
AU - Salomé, R.
AU - Kremer, Y.
AU - Dieudonné, S.
AU - Léger, J. F.
AU - Krichevsky, O.
AU - Wyart, C.
AU - Chatenay, D.
AU - Bourdieu, L.
N1 - Funding Information:
We thank Anne Feltz, Jens Eilers and Luca Guidoni for fruitful discussions. We are grateful to Olivier Crégut for lending us his auto-correlator. This work has been supported by a “Human Frontier Science Foundation” research grant, by the “Ministère de la Recherche et de l’Enseignement Supérieure” (Actions Concertées Incitatives Jeunes Chercheurs 1999), the CNRS (Programme Physique et Chimie du Vivant 1997, Programmes Ultimatech 1997 and 1998), the SPM department of CNRS (Projet jeune chercheur 1999, ATIP jeune chercheur 2002), the French-Israeli Collaboration Program Arc en ciel 2001/02 and the TMR Marie Curie Research Training Grant.
PY - 2006/6/30
Y1 - 2006/6/30
N2 - 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° 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 (1 kHz). 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.
AB - 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° 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 (1 kHz). 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.
KW - Acousto-optical deflectors (AODs)
KW - Calcium imaging
KW - Multiunit recording
KW - Random-access two-photon scanning microscopy (RA-TPSM)
KW - Spatial pre-compensation
KW - Temporal pre-compensation
KW - Two-photon scanning microscopy (TPSM)
KW - Ultrafast scanning
UR - http://www.scopus.com/inward/record.url?scp=33744547575&partnerID=8YFLogxK
U2 - 10.1016/j.jneumeth.2005.12.010
DO - 10.1016/j.jneumeth.2005.12.010
M3 - Article
C2 - 16458361
AN - SCOPUS:33744547575
SN - 0165-0270
VL - 154
SP - 161
EP - 174
JO - Journal of Neuroscience Methods
JF - Journal of Neuroscience Methods
IS - 1-2
ER -