This instrument proposal is for an inverted Zeiss LSM880 Fast Airyscan laser scanning confocal microscope with time-correlated single photon counting (TCSPC) fluorescence lifetime imaging (FLIM) and fluorescence correlation spectroscopy (FCS) capabilities for use in the Cancer Research Laboratory?s Molecular Imaging Center (MIC) at the University of California, Berkeley. UC Berkeley has a strong record of cross-disciplinary discoveries and innovation and this instrument housed in the active, highly-regarded MIC will catalyze breakthroughs for a large group of prominent researchers from five colleges on the campus: the renowned College of Chemistry, the College of Letters and Sciences, the College of Engineering, the College of Natural Resources, and the School of Optometry. The group comprises 18 well- funded NIH, NSF and DOE researchers working on a range of projects of significant biological and health- related interests. The Zeiss LSM880 Fast Airyscan combines a unique 32-channel detector for improved resolution in x, y, and z, with the ability to also shape the beam into a small array for 4 times faster scanning without sacrifice to sensitivity or resolution relative to standard laser scanning confocal microscopy (LSCM). TCSPC FLIM is a very sensitive, quantitative method to determine information about the molecular environment, position and rotation of a fluorophore. Standard fluorescence microscopes do not have the capability to measure the typically nanosecond decay times of standard fluorescent proteins and probes. FLIM is especially useful for measuring angstrom level interactions between fluorophores (Frster resonant energy transfer, FRET). The same instrument will allow us to also measure fluorescence correlation spectroscopy (FCS) which can be used to measure diffusion time and can provide information on concentration and number of molecules. FLIM and FCS, while less well-known than LSCM, are extremely powerful techniques, and even more so when combined with the strengths of a super-resolution laser scanning confocal microscope. While the areas of study for the user group are diverse and overlapping, the need for specialized, high-resolution, time-domain imaging along with high-resolution LSCM unites the group. This instrument will empower efforts by UC Berkeley researchers to develop the tools and techniques to aid in diagnosis, understanding, treatment and prevention of a number of environmental and health- related problems, including climate change, starvation, Alzheimer?s disease, Parkinson?s disease, autism, cardiac arrhythmias, myopia, substance abuse, and cancer.
This instrument proposal is for a fast, super-resolution laser scanning confocal microscope with fluorescence lifetime and fluorescence correlation spectroscopy capabilities. The new instrument will allow researchers to visualize high-resolution, molecular-level information in 4-dimensions (x, y, z, and t) from living cells and tissues. Access to this instrument in the Molecular Imaging Center at the University of California, Berkeley, will enable investigations into the fundamental mechanisms underlying important biological processes including transcription, voltage regulation, touch and force sensation, toxic substance detection, cellular homeostasis, and photosynthesis.
