Ultrafast random-access scanning in two-photon microscopy using acousto-optic deflectors
Ultrafast random-access scanning in two-photon microscopy using acousto-optic deflectors
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DOI：
10.1016/j.jneumeth.2005.12.010
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发表时间：
2006-06-30
影响因子：
3
通讯作者：
Bourdieu, L.
中科院分区：
文献类型：
--
作者：
Salome, R.;Kremer, Y.;Bourdieu, L.
双光子扫描显微镜（TPSM）是一种强大的工具，成像深层组织与亚细胞分辨率。然而，TPSM的时间分辨率受到用于引导激光束的振镜的强烈限制。因此，快速的生理事件只能通过重复扫描视野内的单线来跟踪。由于声光偏转器（aod）是非机械装置，它们允许在微秒时间尺度上进入视场内的任何点，因此是提高TPSM时间分辨率的绝佳候选者。然而，使用基于aod的飞秒脉冲扫描仪会带来一些技术难题。本文描述了一种基于两个交叉aod的全数字TPSM装置。它特别包括一个声光调制器（AOM），放置在相对于aod的45位置，以预先补偿aod中发生的飞秒脉冲的大空间畸变，以优化空间分辨率和荧光激发。我们的设置允许从高速（1千赫）自由选择的兴趣点记录。随机存取TPSM （RA-TPSM）通过最大化在感兴趣点上花费的时间，构成了生物组织中毫秒级分辨率的多单元记录的一种很有前途的方法。(c) 2005 Elsevier B.V.版权所有
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. (c) 2005 Elsevier B.V. All rights reserved.