This proposal requests funds for equipment for fluorescence and video-enhanced microscopy to include a cooled charged coupled device (CCD) camera and imaging system for detecting low intensity fluorescence, a fluorescence microscope equipped with infinity corrected and differential interference contrast optics, an image processing system and Newvicon camera for video-enhanced microscopy, and a micromanipulator and piezoelectric microinjector for use with plant cells. This is a joint proposal by 7 investigator (5 major users, Wendy F. Boss, Rebecca S. Boston, Mark Conkling, Dominique Robertson, Ron Sederoff and Ross Whetten, and 2 minor users, Tim Petty and Kelly Tatchell) from four departments and two colleges. To our knowledge there is no other cooled CCD camera on campus and there is no comparable equipment for high resolution video-enhanced microscopy in the College of Agriculture and Life Sciences or the College of Forestry Resources. The principle investigators study basic cellular processes and signal transduction using a combination of molecular, genetic, biochemical, and microscopic techniques. The research projects include signal transduction (Boss), protein trafficking and packaging to storage vesicles in maize (Boston), root-specific gene expression and nematode resistance (Conkling), viral pathogenesis (Robertson and Petty), cell division and cell cycle control (Tatchell), and lignin biosynthesis, fungal disease resistance and genetic mapping in loblolly pine (Sederoff and Whetten). For the above studies we need to visualize fluorescent proteins within living cells, to monitor changes in protein distribution and cellular morphology over time, and to resolve fluorescent markers on metaphase chromosomes. Our research is currently limited by the sensitivity and resolution of existing microscopic equipment. The Botany Department has made a commitment to the facility by providing space and technical support. The equipment will be under the s upervision of Dr. Wendy F. Boss and Mr. Dana Moxley, and it will be housed in a central location. The requested equipment will contribute significantly to our ongoing efforts to strengthen cell biology at North Carolina State University.
