Award details

Analysis of organelle protein and ion dynamics in plant and fungal cells by blue-excitation META confocal microscopy

Reference	REI20537
Principal Investigator / Supervisor	Professor Ian Moore
Co-Investigators / Co-Supervisors	Professor Mark Fricker, Professor Christopher Hawes, Professor Marc Knight, Professor P Rainey
Institution	University of Oxford
Department	Plant Sciences
Funding type	Research
Value (£)	73,000
Status	Completed
Type	Research Grant
Start date	01/01/2004
End date	31/12/2008
Duration	60 months

Abstract

The funding we request will upgrade our current LSM510 confocal microscope to a state-of-the-art instrument unique in the field of plant and microbial sciences in the UK. Confocal microscopy has become a key method in cell and developmental biology. In combination with the development of new fluorescent probes, confocal microscopy is also being applied increasingly to protein and small molecule biochemistry in living cells. Although our current confocal microscope is a top-of-the-range instrument used extensively by several research groups, its use in some major research programmes is precluded by the lack of suitable laser wavelengths, filter combinations, or autofluorescence problems. Furthermore, in several key areas the principal technical constraint in implementing our experimental strategy is the limitation imposed by the confocal technology available. We will overcome these technical obstacles by adding a 25mW 405nm diode laser and a META spectral deconvolution system. This will improve the technical capability of our microscope in the following key areas: 1. imaging fluorescent dyes with deep blue or near UV excitation peaks; 2. selective excitation of CFP in multiple labelling, co-localisation and FRET studies; 3. photoactivation of PA-GFP in studies of protein and organelle dynamics; 4. simultaneous discrimination of multiple fluorescent proteins and conventional dyes; 5. independent control of fluorophore signal strength using META imaging; 6. compensation for auto-fluorescence of plant and microbial cells. These technical improvements will facilitate the use of GFP-based technologies, thiol dyes, DNA dyes, and cell wall dyes in a wide spectrum of plant and microbial research ranging from bacterial population genetics, to plant pathogen interactions, intracellular membrane traffic, plant thiol metabolism, signalling in response to biotic stress, long distance transport in fungi, plant developmental biology, and biosensors.

Summary

unavailable

Committee	Closed Committee - Plant & Microbial Sciences (PMS)
Research Topics	Microbiology, Plant Science
Research Priority	X – Research Priority information not available
Research Initiative	Research Equipment Initiative 2003 (RE2) [2003]
Funding Scheme	X – not Funded via a specific Funding Scheme

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