A Two Photon AOD Microscope

描述（由申请人提供）：当前的双光子（2P）共聚焦成像系统是围绕传统的共聚焦显微镜扫描头构建的，该扫描头使用振镜来控制激光束扫描。这种技术极大地限制了扫描速度。基于声光偏转器（AOD）的2P显微镜的最新发展，我们将测试开发一种商用的基于AOD的2P成像系统的可行性。成功将允许科学家使用成像速度范围从目前商业系统上可用的到每秒30帧。我们的建议包括4个具体目标。具体目标1：我们将建立一个alpha版本的2P-AOD显微镜，该显微镜将使用超快脉冲激光激励源。我们将对一年前研发的2P显微镜Ultima进行改进，用于AOD扫描，并构建扫描控制和信号调理电路，为高速2P成像提供控制和采集软件。

DESCRIPTION (provided by applicant): Current two-photon (2P) confocal imaging systems are built around traditional confocal microscope scan heads that use galvanometers to control laser beam scanning. This technology significantly limits scanning speed. Building on recent developments of acousto-optical deflector (AOD)-based 2P microscopes we will test the feasibility of developing a commercially available AOD-based 2P imaging system. Success will permit scientists to use imaging speeds ranging from those available on current commercial systems up to 30 frames per second. Our proposal includes 4 specific aims. Specific Aim I: We will build an alpha version 2P-AOD microscope that will operate with an ultra-fast pulse laser excitation source. We will modify a recently developed 2P microscope, the Ultima, which we have developed in the past year, for AOD scanning and will construct scan control and signal conditioning circuitry to provide control and acquisition software for high-speed 2P imaging. Specific Aim II: We will extend the recent development of 'Prairie View' software, galvanometer-based confocal and two-photon (2P) software, to control the 2P-AOD microscope. Specific Aim III: We will compare the optical resolution of the 2P-AOD microscope with that of current galvanometer-scanning 2P systems (a Bio-Rad-based system, a Nikon-based C1 system and the Prairie Ultima). Specific Aim IV: We will test the performance of the 2P-AOD microscope in biological applications. Success in this project will provide the scientific community with an innovative microscope that will enhance its ability to perform live cell imaging in a range of scientific fields, including cell biology, neuroscience and regenerative medicine, and allow innovative studies in such areas as synaptic transmission, learning and memory, and regenerative medicine and stem cell biology.