Funds are requested to purchase a Zeiss LSM MP multiphoton microscope with a Coherent Chameleon Ultra II Ti:Sapphire pulsed laser. This instrument will replace our current BioRad Radiance Multiphoton Microscope, which no longer meets the needs of our users. In addition, Carl Zeiss is phasing out their legacy BioRad systems and no longer stocks replacement hardware or provides software upgrades for the Radiance 2300 system. The dedicated multiphoton microscope will be housed in the Imaging/Physiology Core Facility supported by the NCRR Center of Biomedical Research Excellence (COBRE) in Neuroscience grant (5P20 RR16435, 07/01/06 - 06/30/11). The competitive renewal of the COBRE grant is expected to start 7/1/01/2011 and provide 5 additional years of Core support. The Neuroscience COBRE Imaging/Physiology Core is administered by a full-time, highly trained microscopist. Acquisition of the Zeiss LSM 7 MP dedicated multiphoton microscope, a modern, modular, and actively supported system, would greatly enhance the capabilities of the Core and would meet the emerging needs of the present Core investigators and provide the opportunity for additional funded investigators to significantly expand the scope of their research programs. The new Zeiss multiphoton microscope, model LSM 7MP, offers impressively rapid acquisition speed, about 4 times higher than the speed of our BioRad Radiance 2100 MPD two-photon microscope. The high acquisition speed combined with the superior sensitivity and lower noise levels makes LSM 7 MP perfect for studying the activity of living neurons and other cell types in the brain. In recent years, multiphoton photolysis of caged compounds (uncaging) is increasingly used as a tool to quickly deliver calcium and other substances intracellularly or in the extracellular space with great precision in time, location and volume. Furthermore, multiphoton uncaging allows deeper penetration into live tissues, with smaller volumes compared to single photon photolysis. Our highly innovative research projects that investigate the neuro-vascular coupling in the brain and cerebral arteriole smooth muscle and endothelium function absolutely require the use of photolysis of caged calcium, IP3, glutamate and ATP. The Zeiss LSM 7 MP is capable of two photon photolysis localized within multiple arbitrarily defined regions of interest without slowing down the acquisition speed. This will allow us to simultaneously uncage and image living cells or tissue with a speed that is adequate for monitoring and recording the changes in cellular activity that occur as a result of uncaging. Moreover, this will enable us to simultaneously or consecutively uncage compounds in more than one cell or location thus providing us with the unique opportunity to study the interactions and communications between different cells or structures in the brain.

Public Health Relevance

This instrument would support research designed to understand the development and normal functioning of the brain and autonomic organs and their alteration in response to disease. Imaging of cellular responses within brain slices and organs will provide key insight into the regulation of neuronal, vascular, and connective tissue function and their interdependence. This insight will inform development of therapeutic approaches for diseases that strike these tissues.

National Institute of Health (NIH)
Office of The Director, National Institutes of Health (OD)
Biomedical Research Support Shared Instrumentation Grants (S10)
Project #
Application #
Study Section
Special Emphasis Panel (ZRG1-CB-N (30))
Program Officer
Levy, Abraham
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of Vermont & St Agric College
Anatomy/Cell Biology
Schools of Medicine
United States
Zip Code
Harraz, Osama F; Longden, Thomas A; Dabertrand, Fabrice et al. (2018) Endothelial GqPCR activity controls capillary electrical signaling and brain blood flow through PIP2 depletion. Proc Natl Acad Sci U S A 115:E3569-E3577
Balbi, Matilde; Koide, Masayo; Schwarzmaier, Susanne M et al. (2017) Acute changes in neurovascular reactivity after subarachnoid hemorrhage in vivo. J Cereb Blood Flow Metab 37:178-187
Syed, Arsalan U; Koide, Masayo; Brayden, Joseph E et al. (2017) Tonic regulation of middle meningeal artery diameter by ATP-sensitive potassium channels. J Cereb Blood Flow Metab :271678X17749392
Fuchs, Jason R; Darlington, Shelby W; Green, John T et al. (2017) Cerebellar learning modulates surface expression of a voltage-gated ion channel in cerebellar cortex. Neurobiol Learn Mem 142:252-262
Balbi, Matilde; Koide, Masayo; Wellman, George C et al. (2017) Inversion of neurovascular coupling after subarachnoid hemorrhage in vivo. J Cereb Blood Flow Metab 37:3625-3634
Pappas, Anthony C; Koide, Masayo; Wellman, George C (2016) Purinergic signaling triggers endfoot high-amplitude Ca2+ signals and causes inversion of neurovascular coupling after subarachnoid hemorrhage. J Cereb Blood Flow Metab 36:1901-1912
Pappas, Anthony C; Koide, Masayo; Wellman, George C (2015) Astrocyte Ca2+ Signaling Drives Inversion of Neurovascular Coupling after Subarachnoid Hemorrhage. J Neurosci 35:13375-84
Pappas, Anthony; Wellman, George C (2015) Setting the pace for GI motility: ryanodine receptors and IP3 receptors within interstitial cells of Cajal. Focus on ""Intracellular Ca2+ release from endoplasmic reticulum regulates slow wave currents and pacemaker activity of interstitial cells of Cajal Am J Physiol Cell Physiol 308:C606-7
Koide, Masayo; Wellman, George C (2015) Activation of TRPV4 channels does not mediate inversion of neurovascular coupling after SAH. Acta Neurochir Suppl 120:111-6