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

R. Salomé; Y. Kremer; S. Dieudonné; J. F. Léger; O. Krichevsky; C. Wyart; D. Chatenay; L. Bourdieu

doi:10.1016/j.jneumeth.2005.12.010

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

R. Salomé, Y. Kremer, S. Dieudonné, J. F. Léger, O. Krichevsky, C. Wyart, D. Chatenay, L. Bourdieu

Department of Physics

Research output: Contribution to journal › Article › peer-review

132 Scopus citations

Abstract

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.

Original language	English
Pages (from-to)	161-174
Number of pages	14
Journal	Journal of Neuroscience Methods
Volume	154
Issue number	1-2
DOIs	https://doi.org/10.1016/j.jneumeth.2005.12.010
State	Published - 30 Jun 2006

Keywords

Acousto-optical deflectors (AODs)
Calcium imaging
Multiunit recording
Random-access two-photon scanning microscopy (RA-TPSM)
Spatial pre-compensation
Temporal pre-compensation
Two-photon scanning microscopy (TPSM)
Ultrafast scanning

ASJC Scopus subject areas

General Neuroscience

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10.1016/j.jneumeth.2005.12.010

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@article{f79e0dad5c1240ab9d2d48e9d1a6c77e,

title = "Ultrafast random-access scanning in two-photon microscopy using acousto-optic deflectors",

abstract = "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.",

keywords = "Acousto-optical deflectors (AODs), Calcium imaging, Multiunit recording, Random-access two-photon scanning microscopy (RA-TPSM), Spatial pre-compensation, Temporal pre-compensation, Two-photon scanning microscopy (TPSM), Ultrafast scanning",

author = "R. Salom{\'e} and Y. Kremer and S. Dieudonn{\'e} and L{\'e}ger, {J. F.} and O. Krichevsky and C. Wyart and D. Chatenay and L. Bourdieu",

note = "Funding Information: We thank Anne Feltz, Jens Eilers and Luca Guidoni for fruitful discussions. We are grateful to Olivier Cr{\'e}gut for lending us his auto-correlator. This work has been supported by a “Human Frontier Science Foundation” research grant, by the “Minist{\`e}re de la Recherche et de l{\textquoteright}Enseignement Sup{\'e}rieure” (Actions Concert{\'e}es 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.",

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doi = "10.1016/j.jneumeth.2005.12.010",

language = "English",

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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.

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

Abstract

Keywords

ASJC Scopus subject areas

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