MULTIPHOTON MICROSCOPE FOR PURDUE UNIVERSITY

这种多光子显微镜的应用建立在过去六年在癌症中心和普渡大学校园内开发共聚焦显微镜应用的基础上。这所大学目前还没有多光子显微镜。在过去的几年里，稳定数量的要求进行实验，超过了目前的共聚焦能力，要求我们考虑一个更先进的技术。两个具体的实验领域为我们目前的共聚焦显微镜系统造成了重大困难：在动力学协议中的活细胞系统的深层组织成像和分析。深层组织成像成为一个经常性的主题，要求使用共聚焦，并最终得出结论，我们已经达到了我们目前的仪器的能力。我们目前的文书仍然是一项非常重要的文书。事实上，这是一个巨大的资源，我们期望继续使用这一工具。具体来说，它已成为反射成像的首选仪器，并将继续在校园的生物成像计划中发挥重要作用。我们的共聚焦用户面临的第二个困难是活细胞的长期成像。这些测定包括功能测定、分裂或分化细胞的观察、深部组织细胞的表型分析和包括钙通量的细胞功能分析。活细胞研究的需求很大，用我们目前的系统很难充分进行。所要求的仪器将为普渡大学带来它目前不具备的能力。从最初考虑我们需要多光子开始，我们已经等了将近3年。虽然我们在三年前就展示了它的用途，但我们认为当时的技术还太不成熟。在我们的环境中，目前几乎是交钥匙的系统更受欢迎。这项应用的资金将为我们目前的共聚焦能力带来重大进步。

This application for a multiphoton microscope builds upon the past six years of developing confocal microscopy applications in the Cancer Center and across the Purdue University campus. There are currently no multiphoton microscopes at this university. Over the past few years, a steady number of requests to perform experiments that exceeded the current confocal capability, required us to consider a more advanced technology. Two specific areas of experimentation created significant difficulties for our current confocal microscope system: deep tissue imaging and analysis of live cell systems in a kinetic protocol. The deep tissue imaging became a recurrent theme of the requests to use the confocal and culminated in the conclusion that we had reached the abilities of our current instrument. Our current instrument is still a very important instrument. Indeed, it has been a tremendous resource and we anticipate a continuation of the use of this instrument. Specifically it has become the instrument of choice for reflection imaging and will continue to play a significant role in the campus's biological imaging program. The second area of difficulty faced by our confocal users was the long term imaging of live cells. These assays include functional assays, observations of dividing or differentiation cells, phenotyping deep tissue cells, and functional analysis of cells including calcium flux. Live cell studies are in strong demand and will be difficult to perform adequately with our present system. The instrument requested will bring to Purdue University a capability it does not presently possess. We have waited for nearly 3 years, since originally considering our need for multiphoton. While we had demonstrated uses three years ago, it was our opinion that the technology was then too immature. Present-day almost turn-key systems are more desirable in our environment. Funding of this application will bring a significant advance to our present confocal capabilities.