Exploring the physiology and dynamics of plant cells using fast lifetime contrast imaging
Lead Research Organisation:
University of Nottingham
Department Name: Sch of Biosciences
Abstract
We are applying for funding for a cutting-edge Leica Stellaris 8 confocal microscope, equipped with Fluorescence Lifetime Contrast (FALCON) and Digital Light Sheet (DLS) modules. This will replace an aging Leica SP5, a workhorse that has been pivotal in generating over 200 research publications over a period of 15 years, and currently serves 36 regular users. Beyond mere replacement, our proposal introduces novel capabilities that resonate with the evolving landscape of Plant Science research and enable us to pursue areas of BBSRC strategic priority, such as climate-proofing agriculture.
The FALCON module (Fast Lifetime Contrast) is a game-changer, not just measuring signal intensity but also capturing the time a fluorophore retains laser energy, enabling Fluorescent Lifetime Imaging (FLIM). This technology unlocks new avenues, from precise quantification of protein-protein interactions, to quantifying cellular pH and cell membrane tension and separating overlapping fluorescent signals. Example projects where we will use these capabilities include: investigating the wave of signals that trigger the release of plant signals (such as the hormone ABA) during drought, assessing photosynthetic efficiency under future climates, and understanding the mechanical processes by which roots navigate soil to acquire both water and nutrients.
Currently there are only a few microscopes with FALCON capabilities in the UK. We have specified what we believe to be the first microscope in the UK combining FALCON with Digital Light Sheet (DLS). The DLS module further enhances our microscope's versatility, enabling seamless switching between light sheet and confocal/FALCON acquisition modes. This dynamic capability empowers us to conduct fast 3D acquisitions of large samples and to target specific regions with higher Z resolution, providing a comprehensive view from cellular to organ scales. The only other microscope which we know of with similar configuration is in the University of Helsinki, Finland. We already work closely with labs there, and will continue to do so to develop shared methodology. These capabilities will be unique in the UK and dramatically enhance our ability to address crucial questions in cell biology, whilst framing our results within a whole organ/organism context.
Our central location in the Midlands allows access for researchers from nearby universities, such as Warwick and Sheffield. Our goal is to provide accessibility for a regional user base and foster collaboration among plant scientists. One of our strengths as a university is that our expertise extends beyond image acquisition, to include downstream processes, such as image analysis and incorporation of imaging data into mathematical models of plant development. Together with colleagues in Maths and Computer science, we will apply artificial intelligence to analyse the extra information that fluorescence lifetime gives over traditional imaging datasets.
This instrument not only addresses our immediate research needs by replacing a well used and ageing machine, but propels Plant Science into a new era, promising groundbreaking discoveries and collaborations in the area of climate proofing agriculture with far-reaching implications for regional and national research initiatives.
The FALCON module (Fast Lifetime Contrast) is a game-changer, not just measuring signal intensity but also capturing the time a fluorophore retains laser energy, enabling Fluorescent Lifetime Imaging (FLIM). This technology unlocks new avenues, from precise quantification of protein-protein interactions, to quantifying cellular pH and cell membrane tension and separating overlapping fluorescent signals. Example projects where we will use these capabilities include: investigating the wave of signals that trigger the release of plant signals (such as the hormone ABA) during drought, assessing photosynthetic efficiency under future climates, and understanding the mechanical processes by which roots navigate soil to acquire both water and nutrients.
Currently there are only a few microscopes with FALCON capabilities in the UK. We have specified what we believe to be the first microscope in the UK combining FALCON with Digital Light Sheet (DLS). The DLS module further enhances our microscope's versatility, enabling seamless switching between light sheet and confocal/FALCON acquisition modes. This dynamic capability empowers us to conduct fast 3D acquisitions of large samples and to target specific regions with higher Z resolution, providing a comprehensive view from cellular to organ scales. The only other microscope which we know of with similar configuration is in the University of Helsinki, Finland. We already work closely with labs there, and will continue to do so to develop shared methodology. These capabilities will be unique in the UK and dramatically enhance our ability to address crucial questions in cell biology, whilst framing our results within a whole organ/organism context.
Our central location in the Midlands allows access for researchers from nearby universities, such as Warwick and Sheffield. Our goal is to provide accessibility for a regional user base and foster collaboration among plant scientists. One of our strengths as a university is that our expertise extends beyond image acquisition, to include downstream processes, such as image analysis and incorporation of imaging data into mathematical models of plant development. Together with colleagues in Maths and Computer science, we will apply artificial intelligence to analyse the extra information that fluorescence lifetime gives over traditional imaging datasets.
This instrument not only addresses our immediate research needs by replacing a well used and ageing machine, but propels Plant Science into a new era, promising groundbreaking discoveries and collaborations in the area of climate proofing agriculture with far-reaching implications for regional and national research initiatives.
