BLI NEWS
SHARED INSTRUMENTATION GRANT AWARDED
Associate Professor Vickie LaMorte, Ph.D., and her co-PI, Tatiana Krasieva, Ph.D., have been awarded a $500,000 Shared Instrumentation Grant from the National Institutes of Health to purchase a new Zeiss LSM 510 META NLO microscope. Additional funds for the remaining cost of the $750,000 system were arranged by Dr. LaMorte through Beckman Laser Institute and the Department of Biomedical Engineering.
The new Zeiss LSM 510 META NLO laser-scanning microscope is a state-of the art system. Housed on the platform of a Zeiss inverted Axiovert 200 M microscope, it combines standard fluorescence confocal imaging at six different excitation wavelengths with multi-photon fluorescence/second harmonic generation. The excitation source for this modality is a novel femto-second Chameleon Ultra laser (Coherent Inc.), which is tunable at an extremely wide range of wavelengths: from 690 to 1040 nm. One of the unique features of this system is its ability to detect second harmonic generation signals in both transmitted and reflected geometry. META detection provides fast, reliable and precise separation of the emission signals for multiple-labeled samples with widely or almost completely overlapping emission spectra of fluorophores on a pixel-by pixel basis. Specifically, the new Zeiss META imaging system with two photon capabilities will enhance Dr. LaMorte's ability to examine protein-protein interactions occurring within a single cell. Coupled with Fluorescence Resonance Energy Transfer or FRET-based methodologies, this system will provide spectral separation of the GFP molecules that Dr. LaMorte and her lab use to tag the proteins they are interested in examining with pixel-by-pixel resolution. In combination with the subcellular imaging volume that can be achieved by two photon excitation, they will have resolution of these cellular events that was not obtainable with the previous existing systems. Not only will Dr. LaMorte and her lab be able to determine if two proteins interact in vivo within the confines of the cell but now they will be able to see precisely where this interaction is occurring within the cell's architecture. Ultimately, by mapping a protein's dynamic interplay with other proteins in the cell, a fundamental understanding of how this protein contributes to normal and diseased states will be achieved.
The system will be housed and supported within the well-established NIH Biomedical Technology Resource Center (Laser Microbeam and Medical Program, LAMMP) at the Beckman Laser Institute and will be used by a group of 13 multi-disciplinary investigators representing ten departments at the University of California, Irvine. The LAMMP facility is dedicated to defining and understanding broad issues related to the use of light as a tool to selectively perturb, monitor, and image physiology in cells and tissues. According to Dr. Krasieva, manager of the LAMMP facility, all of the original proposed projects were initiated as pilot/feasibility studies using a multi-photon excitation laser scanning system that she and her colleagues built themselves and/or a 14-year-old LSM 410 confocal system. Now that the original projects have expanded in scope and require frequent and often heavy use of laser scanning microscopy, there was an urgent need for a commercial-grade, user-friendly confocal/multi-photon microscope to be housed and managed within the facility. The Zeiss META instrument will ideally accommodate the researchers' needs, which range from low-light-level endogenous signal detection of both fluorescence and second harmonic generation (SHG) in tissues to cellular FRET-based imaging.
Dr. LaMorte expects that the Zeiss META instrument will have a significant overall benefit to the UCI research community and beyond based on the existing strong impact of the LAMMP and its dissemination of new discoveries to colleagues and collaborators.