A Two Photon AOD Microscope

描述（由申请人提供）：当前的双光子（2 P）共焦成像系统是围绕传统的共焦显微镜扫描头构建的，该扫描头使用检流计来控制激光束扫描。这项技术大大限制了扫描速度。基于声光偏转器（AOD）的2 P显微镜的最新发展的基础上，我们将测试开发一个商用的基于AOD的2 P成像系统的可行性。成功将使科学家能够使用成像速度，从目前的商业系统上可用的速度到每秒30帧。我们的建议包括四个具体目标。具体目标一：我们将建立一个阿尔法版本的2 P-AOD显微镜，将与超快脉冲激光激发源。我们将修改最近开发的2 P显微镜，Ultima，我们在过去一年中开发的，用于AOD扫描，并将构建扫描控制和信号调理电路，以提供高速2 P成像的控制和采集软件。 具体目标二：我们将扩展最近开发的“Prairie View”软件，基于检流计的共聚焦和双光子（2 P）软件，以控制2 P-AOD显微镜。具体目标三：我们将比较2 P-AOD显微镜的光学分辨率与当前的振镜扫描2 P系统（基于Bio-Rad的系统，基于尼康的C1系统和Prairie Ultima）。具体目标IV：我们将测试2 P-AOD显微镜在生物应用中的性能。该项目的成功将为科学界提供一种创新的显微镜，提高其在细胞生物学、神经科学和再生医学等一系列科学领域进行活细胞成像的能力，并允许在突触传递、学习和记忆、再生医学和干细胞生物学等领域进行创新研究。

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.