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，大量的实验要求超出了当前的共聚焦能力，这就要求我们考虑一种更先进的技术。两个特定的实验领域为我们目前的共聚焦显微镜系统创造了重大的困难：深层组织成像和活细胞系统的动力学分析。深层组织成像成为反复要求使用共聚焦的主题，并最终得出结论，我们已经达到了我们目前的仪器的能力。我们现在的仪器仍然是一个非常重要的仪器。的确，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，它已经成为反射成像的首选仪器，并将继续在校园生物成像项目中发挥重要作用。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、分裂或分化细胞的观察、深层组织细胞的表型分析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，很难用我们目前的系统进行充分的研究。所要求的仪器将给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演示了使用，但我们认为当时的技术还太不成熟。在我们的环境中，当今几乎是交钥匙式的系统更为可取。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.