MRI: Development of a scan-less temporal focusing two-photon fluorescence microscope for high-speed three-dimensional imaging

得克萨斯大学埃尔帕索分校(UTEP)被授予开发一种高速光学显微镜，用于成像荧光物体并同时跟踪它们在三维(3D)空间中的快速运动。这个跨学科项目通过提供生物、化学、物理和电气工程领域的前沿研究经验，丰富了本科生和研究生的教育。它还培养博士后研究人员，他们将形成下一代乐器演奏家。该项目为生物和化学科学家提供了开发和使用创新和当代仪器的强大背景，显著增强了他们进行尖端研究的能力。开发的显微镜将作为生物光子学核心设施的基石，该设施将促进合作研究、教育和外联活动。西班牙裔占多数(近80%)的UTEP的少数民族学生和地区用户将被邀请参加使用这一设施的研讨会和研究活动。该项目是荧光显微镜领域的一项创新。这种新型显微镜集成了时间聚焦和脉冲整形两种超快激光技术，具有前所未有的高速获取三维体图像的能力(50体积/S，体积尺寸100um×100um×100um)。该显微镜消除了当前用于共焦和双光子显微镜的激光扫描机制，并集成了脉冲整形技术，增加了深度扫描能力。该项目还将开发一种计算机化图像处理系统，该系统将自动分析高速视频。这些能力，即高速和3D体积成像，将使研究人员能够探索和分析以前无法从这种自动图像分析系统获得的快速动态信息。最初的应用领域是海洋病毒感染、纳米颗粒运输和苍蝇脑成像。应用可以扩展到许多其他领域，在这些领域，使用当前的激光扫描荧光显微镜很难研究快速的3D动态过程。

An award is made to the University of Texas at El Paso (UTEP) to develop a high-speed optical microscope for imaging fluorescent objects and simultaneously tracking their fast motion in three-dimensional (3D) space. This interdisciplinary project enriches undergraduate and graduate students' education by providing frontier research experience in the fields of biology, chemistry, physics, and electrical engineering. It also trains postdoctoral researchers who will form the next generation of instrumentalists. This project gives scientists in biology and chemistry a strong background in the development and use of innovative and contemporaryinstruments, significantly enhancing their ability to conduct cutting-edge research. The developed microscope will serve as the cornerstone of a Biophotonics core facility that will promote collaborative research, education and outreach activities. Underrepresented minority students at UTEP, a Hispanic-majority (almost 80%) institution, and regional users will be invited to participate in workshops and research activities using this facility.This project is an innovation in the field of fluorescence microscopy. By integrating two ultrafast laser techniques, temporal focusing and pulse shaping, this new microscope has the capability of acquiring 3D volumetric images at an unprecedented high-speed (50 volumes/s, volume size 100um × 100um × 100um). This microscope eliminates the current laser beam scanning mechanisms typically used in confocal and two-photon microscopes with temporal focusing, and adds depth scanning capability by integrating the pulse shaping technique. This project will also develop a computerized image processing system that will automatically analyze high-speed videos. These capabilities, i.e. high-speed and 3D volumetric imaging, will enable researchers to explore and analyze previously unobtainable fast dynamic information from this automatic image analysis systems. Initial applications are in the areas of marine virus infection, nanoparticle transportation, and fly brain imaging. Applications could extend to many other fields where it is hard to study fast 3D dynamic processes using current laser beam scanning fluorescence microscopes.