Spinning Disk Confocal Microscope System

Providence College, Providence RI

Investigators

Abstract

Project Summary/Abstract Our NIH user group from an Emerging Research Institution requests funds from the NIH BIG program to purchase a Nikon CSU-W1 spinning disk confocal microscope system to be housed at Providence College (PC) in Providence, RI. This acquisition is essential to our NIH-funded investigators who require high- resolution 3D fluorescence microscopy for their projects. Currently, most of our investigators rely on a 14-year- old Zeiss LSM 700 confocal microscope system that is experiencing hardware breakages and software crashes related to its age, and the system has been discontinued from service by Zeiss. This system has major imaging issues including weak laser power, low sensitivity, slow scan speed, photobleaching, and phototoxicity. These issues are damaging to our research programs and limit our ability to perform our NIH-funded research projects. PC has recently completed massive renovations costing over $55 million dollars to create a state-of-the-art Science Complex. Part of this renovation included building a brand-new Microscope and Imaging Facility. The expansion of the Science Complex has been accompanied by the recruitment of new faculty with diverse biomedical research projects with distinct confocal imaging needs. We have a significant need for an upgraded confocal imaging system like the CSU-W1 that allows for diverse imaging applications, including 3D imaging of live or/and fixed cells and tissues. The Nikon CSU-W1 spinning disk confocal is a high-speed, high-resolution imaging system featuring an ultra-wide field of view and enhanced signal-to-noise ratio. The CSU-W1 has a wider inter-pinhole spacing in the spinning disk that reduces pinhole crosstalk allowing deep imaging applications from 50 to 100um. Our CSU-W1 will be equipped with five CFI Plan Apo objectives (4x, 20x, 40x, 60x oil, and 100x oil) and four laser lines (405, 488, 561, and 640nm) that will meet the needs of all users. The Ti2-E inverted microscope base is fully motorized, with a stable, drift-free platform. The Prime BSI Express CMOS camera (2048x2048) has a near perfect 95% quantum efficiency to maximize signal detection. The CSU-W1 also includes a workstation equipped with NIS-elements AR software that allows control of the CSU-W1 imaging system and has built-in multi-X, Y, Z, Time, multichannel experiment capabilities. This instrument allows time-lapse imaging of live cells without complications of phototoxicity, image stitching of large tissue sections, fast acquisition of 3D volumetric multi- channel images, and high-resolution imaging to view fine structures in cells. These features will be used for diverse biomedical research projects to identify structural changes in neurons associated with disease and behavior, cell size and cell division mechanics, host-pathogen interactions, transport processes in squid photoreceptors, cellular effects induced by novel small molecule probes, and signaling pathways induced by microneedles in human skin. This user group has an immediate need for the CSU-W1 spinning disk confocal to support current projects and to expand and innovate their biomedical research programs.