Resource for Multiphoton Characterization of Genetically-Encoded Probes

双光子激光扫描显微镜为许多项目提供动力。大脑倡议 因为这是捕捉活体神经网络协同活动的最佳方式。 用荧光蛋白构建的遗传编码活性探针和传感器是 标记和记录活神经元回路的首选工具。但要 迄今为止，几乎所有这些探针的优化都仅在单光子范围内。 这主要是因为测量定量的双光子吸收特性 需要更先进的设备和专业知识在电子物理和非线性 光学.在过去的五年里，我们的小组非正式地提供了这项服务， 蛋白质工程师和神经科学家：他们给我们新开发的蛋白质， 仔细表征其2 PA特性。虽然这在过去只在一个 尽管只有少数实验室，但BRAIN计划已经推动了更多需要这一点的实验室 我们现在需要支持来建立一个资源。 在这个项目中，我们将建立一个独特的多光子表征资源， 遗传编码探针在蒙大拿州立大学校园（波兹曼，MT） 该公司将继续提供2 PA特性的表征服务， 许多蛋白质工程师和神经科学家都在研究多光子稳定性。 我们计划为调查人员提供非常广泛的访问我们设施的机会， 研究与NIH/NINDS的使命和BRAIN倡议的目标相关。 所获得的信息将通过科学出版物广泛传播， 显微镜网站上。我们将组织年度研讨会， 关于2 PA物理学和测量方法的培训，并提供所有必要的 为那些愿意在实验室开始这种测量的研究人员提供信息。

Two-photon laser scanning microscopy powers many projects in the. BRAIN initiative because it is the best way to capture the concerted activity of living neural networks. Genetically-encoded activity probes and sensors built with fluorescent proteins are the tools of choice for labeling and recording from living circuits of neurons. However, to date, virtually all optimization of these probes has been solely in the one-photon regime. This is mainly because measuring quantitative two-photon absorption properties demands more sophisticated equipment and expertise in photophysics and nonlinear optics. Over the past five years, our group has informally provided this service for protein engineers and neuroscientists: they sent us newly developed proteins and we carefully characterized their 2PA properties. While this has worked in the past for just a few laboratories, the BRAIN initiative has fueled many more laboratories in need of this service to the extent that we now need support to establish a Resource. In this project, we will establish a unique Resource for Multiphoton Characterization of Genetically-Encoded Probes at the Montana State University Campus (Bozeman, MT) that will continue providing the service of characterization of 2PA properties and multiphoton stability to a much large number of protein engineers and neuroscientists. We are planning to provide a very broad access to our facility for Investigators whose research is relevant to the NIH/NINDS mission and to the goals of the BRAIN initiative. The information obtained will be broadly disseminated via scientific publications and one of the microscopy websites. We will organize annual workshops where we will provide training on 2PA photophysics and methods of measurements and provide all necessary information for researchers who were willing to start such measurements in their labs.