PlatformsCellular Neurophysiology

Access to infrastructure and expertise in electrophysiology and imaging at the single cell level.

Click Here for the Cellular Neurophysiology Platform Leaders Report 2003/2004

The core aim of this platform is to build and maintain world-class infrastructure for cellular neurophysiology, primarily in the area of electrophysiology and physiological cell imaging. This facility enables research scientists to access electrophysiology services, including technology, computer engineering, fabrication and specialist support, which have previously been available to only a select expert few. Professor John Furness and Dr Ernie Jennings, who are both based at the Department of Anatomy & Cell Biology of the University of Melbourne, provide scientific leadership. The Platform manager is Mr John Harvey. Based in Victoria, these services are available to the wider national scientific community.

Objectives
Provision of high quality electrophysiological and cell imaging support to core projects and additional assistance to laboratories where electrophysiology or imaging is not their expertise.
Service to laboratories that do not have expert knowledge in electrophysiology by the provision of sophisticated analytic assistance and specialised equipment that can be prohibitively high in cost for those funded through individual research grants.
Ongoing development of cell physiology techniques, including combined imaging and electrophysiology and combined PCR and electrophysiology.

Facilities and Expertise
Core facilities within the Cellular Neurophysiology Platform comprise equipment and personnel at the following locations:
Department of Anatomy and Cell Biology
  8 electrophysiology work stations, which are equipped for intracellular and patch recording, including single-electrode voltage clamp, and which have additional equipment, such as spritz application of drug, nerve stimulators (on all apparatus), infrared DIC and fluorescence illumination. These set-ups include both inverted and upright microscopes and facilities for brain-slice recording.
  Two Biorad 1024 confocal imaging systems on Zeiss microscopes and one conventional multi-wavelength fluorescence microscope with video image capture.
  Laboratory for processing, imaging and analysis of cells after intracellular dye filling.
  Ancillary equipment includes two Brown-Flaming microelectrode pullers, a Brown-Flaming laser puller, micro-osmometer, microforge, Bessel low noise filter and a Zero Z vibratome. There is a small workshop area in the electrophysiology suite for minor electrical and mechanical work (for example, making stimulating electrodes), a small machine shop and electronics workshop, and an ultra-sterile facility for creation and holding of transfected cells.

Department of Physiology, The University of Melbourne and Howard Florey Institute
  Six microelectrode set-ups, two of which are set up for simultaneous Ca2+ or voltage-sensitive dye imaging using confocal microscopy, and another with infrared DIC illumination. Functional imaging systems are set up for both high spatial and high temporal resolution. The facility includes a multi-photon imaging system that is set up for cortical imaging and electrophysiology in vivo.
  Ancillary equipment includes microelectrode pullers (conventional and laser), vibroslicers, stereotactic apparatus for mouse work; three additional Ca2+ imaging set-ups; facility for transfection of frog oocytes and cell lines, and for recording in transfected cells; set up for whole cell patch recording from cortical cells in living mice; and a Xenopus colony.

Department of Physiology, Monash University
  Six electrophysiology workstations, which are equipped for sharp electrode and patch recording. Two of these workstations are set up for single-electrode voltage clamp. Calcium or dye imaging, together with electrophysiology is available on three workstations whilst flash photolysis is available on one workstation. Also, one workstation is equipped with infrared differential interference contrast optics for brain slice electrophysiology. The equipment for the maintenance of and recording from transfected cells lines is also available.
  Ancillary equipment includes: microelectrode pullers (5), patch pipette polisher (2), patch perfusion system (1), vibroslicer (2), picospritzers (3), and stimulators. Fluorescent imaging of living tissues and various aspects of confocal microscopy (3D, 4D multi-channel and co-localisation, FRAP, FRET and volume rendering) of fixed tissues are also available via Monash Micro Imaging.

Prince Henry's Institute of Medical Research, Monash University
  Two patch clamp workstations are fully equipped, with one also equipped for microfluorescent spectra-analysis system for simultaneous Ca2+ measurement and the other equipped with amperometry recording and capacitance recording. An Olympus FV 300 confocal microscope system is also fully functional.

Key Personnel
At the Departments of Anatomy & Cell Biology and Physiology at The University of Melbourne, and the Howard Florey Institute, the scientists who lead the platform are:
Professor John Furness, whose expertise is in intracellular and patch recording from neurons of peripheral ganglia; in the morphological and immunohistochemical analysis of cells following elecrophysiological characterisation; and in studies of proteing trafficking by confocal microscopy.
Dr Ernie Jennings, whose expertise is in recording from brain stem and in pharmacological analysis at the single cell level using electrophysiological techniques.
Dr Steve Petrou, whose expertise is in cell transfection and analysis of the effects of channel (and other) mutations on the electrophysiological properties of neurons. Dr Phil Davies is in the same laboratory.
Dr Chris Reid, whose expertise is in studies of dendritic integration, and in high resolution Ca2+ imaging.
Dr Damian Myers, whose expertise is confocal and multiphoton imaging
Professor David Williams, whose expertise is in combined electrophysiological and imaging studies, particularly Ca2+ imaging in live cells and the study of Ca2+ microdomains. Dr Chris Reid is in the same laboratory.
Professor Bert Sakmann (Max Planck University for Medical Research, Heidelberg, Germany) is a regular visitor and user of the Platform Facilities.
Mr John Harvey, Platform Manager.

At the Department of Physiology and Prince Henry's Medical Research Institute at Monash University, the scientists who lead the platform are:
Dr Helena Parkington, leads a laboratory that is internationally acclaimed for electrophysiological prowess, particularly in relation to relating events occurring in the single channel level back to the whole animal. Together with Drs Harry Coleman and Marianne Tare, this laboratory has expertise in recording from brain slices and peripheral tissues and in imaging.
Dr Chen Chen, expertise in ion channels in neuroendocrine cells and pituitary cell electrophysiology, with broad experience in patch clamp and calcium analysis.
Dr Rick Lang, expertise in sharp electrode and patch recording from peripheral tissues. Dr Betty Exintaris is a member of this laboratory.

At the School of Medical Science at RMIT University:


Dr Martin Stebbing, whose expertise is in recording from brain and spinal cord slices, dissociated cells and peripheral ganglia using sharp and patch electrodes.


The capabilities of the Platform are further enhanced by post-doctoral scientists, research assistants and computer technicians with extensive experience in this field.

Key Achievements 02-03
Currently, the Platform is providing support for the NSV Epilepsy, Neurodegeneration and Pain Programs. These programs would not be able to achieve their aims without Platform support. It is anticipated that scientists within the Stroke program will commence access in the near future.
The Platform is servicing external users from the University of Melbourne, Research Institutes and Industry.
The scientists working within the platform also deal with many casual inquiries and requests for assistance. Within the Platform is a gathering of the leaders in Cellular Neurophysiology currently active in Melbourne. It has an excellent equipment base, and sufficient capacity to cope with several external users at one time.
Professor John B Furness (Centre for Neuroscience & Dept of Anatomy & Cell Biology) was awarded a Centenary Medal for his work in neuroscience and a Distinguished Achievement Award of the Australian Neuroscience Society in 2003.
Seventeen papers that used Platform facilities were published in 2003.
Income from external use of the platform was $88,667 in the financial year 2003-2004.


 
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