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.
