MULTIPHOTON MICROSCOPE FOR PURDUE UNIVERSITY

多光子显微镜的这一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焦显微镜应用的基础上。这所大学目前没有多光子显微镜。在过去的几年里，稳定数量的实验请求超过了目前的共焦能力，这要求我们考虑一种更先进的技术。两个特定的实验领域给我们目前的共聚焦显微镜系统带来了巨大的困难：深层组织成像和以动力学方案分析活细胞系统。深层组织成像成为要求使用共焦的一个反复出现的主题，并最终得出结论，我们已经达到了目前仪器的能力。我们目前的工具仍然是一个非常重要的工具。事实上，这是一项巨大的资源，我们期待着继续使用这一文书。具体地说，它已经成为反射成像的首选工具，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领域是活细胞的长期成像。这些分析包括功能分析、细胞分裂或分化的观察、深层组织细胞的表型，以及包括钙通量在内的细胞功能分析。活细胞研究的需求很大，在我们目前的系统下很难充分发挥作用。所要求的仪器将为普渡大学带来它目前不具备的能力。自从最初考虑我们对多光子的需求以来，我们已经等待了近3年。虽然我们在三年前就已经演示了用途，但我们认为当时的技术太不成熟了。在我们的环境中，今天几乎是交钥匙系统是更可取的。这项申请的资金将为我们目前的共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.