MRI: Development of a Multi-Photon Microscope with Adaptive Optics

加州大学圣克鲁斯分校获得一项奖励，将长波激发光（1,550 nm）和自适应光学（AO）添加到商用多光子显微镜（Olympus）中，用于活体深层组织生物成像。自适应光学通过校正大气造成的模糊，为大型地面望远镜提供了衍射受限的图像。他们建议在多光子显微镜中重新应用类似的自适应光学技术，以获得活体生物组织深处的衍射限制图像。这一发展将为生物学家提供新的成像能力，类似于自适应光学为天文学家提供的优势，使他们能够在神经生物学中获得最高分辨率的深层组织图像，以研究神经系统中的大脑发育和突触可塑性，在细胞生物学中研究细胞周期事件，在发育生物学中研究干细胞，染色质重塑和细胞骨架。该项目的研究活动包括为本科生和研究生提供参与由光学物理学家、电气工程师和生物学家组成的多学科研究团队的机会，为他们培养下一代仪器学家和高级用户做出贡献。研究活动和仪器将通过加州大学圣地亚哥分校的工程和科学课程的实践项目，西班牙裔服务机构（HSI），以及通过国家科学基金会赞助的外展计划，加州州立数学和科学暑期学校（COSMOS），一个为在数学和科学方面表现出兴趣和成就的高中学者提供的暑期住宿计划，整合到本科和研究生水平的教学中。他们还将与加州大学所有十个校区和三个附属国家实验室（LANL， LBNL和LLNL）以及工业界合作，广泛地向生物研究界传播这项技术。这些改进将极大地促进我们对细胞和亚细胞水平上生命过程的基本理解，从而造福社会。所提出的活动是衍射极限成像（738 nm）深度（1500 nm）的两倍多，在动态活组织（AO帧率1 Hz）中发生许多基本的细胞过程，如神经元生长，组织和突触形成。例如，通过将成像深度从500微米增加到1000微米，他们可以到达更深的皮层，以观察发育过程中突触重组和病理条件下突触重组是否遵循与活体动物突触不断重塑的皮层表层相似的规则。通过将成像深度增加到1000米以上，它们可以到达海马体，即学习和记忆的部位。这将有助于研究突触在学习过程中是如何重组的，以及它们是如何编码形成持久记忆的。

An award is made to the University of California Santa Cruz to add long-wavelength excitation light (1,550 nm) and adaptive optics (AO) into a commercial multiphoton microscope (Olympus) for live deeptissue biological imaging. Adaptive optics has provided diffraction-limited images for large ground-based telescopes by correcting for the blurring caused by the atmosphere. They propose to re-apply similar adaptive optics techniques in multiphoton microscopy to obtain diffraction-limited images from deep within living biological tissues. This development will provide biologists with new imaging capabilities, similar to advantages adaptive optics provided to astronomers, enabling them to obtain the highestresolution deep-tissue images in neurobiology to study brain development and synapse plasticity in the nervous system, in cell biology to study cell cycle events, and in developmental biology to study stem cells, chromatin remodeling and the cytoskeleton. The research activities in this project include opportunities for undergraduate and graduate students to participate in a multidisciplinary research team consisting of optical physicists, electrical engineers, and biologists, contributing to their training as the next generation of instrumentalists and advanced users. The research activities and instrumentation will be integrated into teaching at the undergraduate and graduate levels through hands-on projects in engineering and science courses at UCSC, a Hispanic Serving Institution (HSI), and through an NSF sponsored outreach program, the California State Summer School for Mathematics and Science(COSMOS), a summer residential program for high school scholars with demonstrated interest and achievement in math and science. They will also work in a partnership with all ten UC campuses and three affiliated national labs (LANL, LBNL, and LLNL) and industry to broadly disseminate the technology to the biological research community. Such improvements will benefit society by greatly advancing our fundamental understanding of life processes at the cellular and sub-cellular levels.The proposed activity more than doubles the depth (1,500nm) of diffraction limited imaging (738 nm) into dynamic live tissue (AO frame rate 1 Hz) where many fundamental cellular processes occur, such as neuron growth, organization and synapse formation. For example, by increasing the imaging depth from 500 um to 1,000 um, they can reach the deeper cortical layers to see if synaptic reorganization during development and under pathological conditions follows similar rules as in the superficial cortical layers, were synapses are constantly remodeling in living animals. By increasing the imaging depth beyond 1,000 um they can reach the Hippocampus, the site for learning and memory. This will allow studies of how synapses reorganize during learning and how they encode for long-lasting memory.