Maestro Pro multiwell microelectrode array for the University of Liverpool electrophysiology suite: Cell physiology meets high throughput.
Lead Research Organisation:
University of Liverpool
Department Name: Musculoskeletal & Ageing Science
Abstract
This proposal is for the Institute of Life Course and Medical Sciences (ILCAMS), in the University of Liverpool to purchase a new equipment that can perform physiological recording on a much larger scale, and with greater speed than that which was possible in the past.
*What are we requesting?*
Much of modern biology is based around identifying large numbers of biomolecules in cells and tissues and the pathways that they influence. Due to extraordinary breakthroughs over recent years this can now be done at astonishing speeds unthinkable in the past. For example single cell RNA-sequencing can now identify almost the entire mRNA complement (~active genes) from 50,000 individual cells in one day (may take a couple of days to analyse the data!). However, one area of biology that is still lagging far behind is physiological recording; recording what changes occur in cell electrical behaviour, movement, survival, contraction etc. These are generally performed by highly skilled experts, but typically at the rate of about one cell per day; once trained and all set up. What we are requesting is a new generation of physiological recording apparatus that can identify physiological changes in hundreds of cells per day. Not up to the speed yet of single cell RNA-sequencing, but a massive advance on what was previously possible for us, or any other labs in the North West of England.
*What can this equipment do that existing equipment cannot?*
Whilst this apparatus would be useful across a range of disciplines from genetics, cell biology neuroscience, ageing, stem cells etc, there are three specific facets we need to highlight.
(1) Existing equipment records only one cell at a time or one recording from many cells all aggregated together. That is painfully slow. This new machine allows for the recording of dozens of cells simultaneously making high throughput electrical and contractility assays possible.
(2) As an extension of (1) this machine will allow one to record individual cells not just faster, but in parallel in a structured fashion so that one can track network activity in real time. Cells within tissues do not behave as individual mavericks doing their own thing; they interact in networks and this machine allows these networks to be analysed in Liverpool for the first time.
(3) This equipment allows analysis of cell integrity, movement and electrical properties all at the same time across dozens of cells in the network and in several parallel experiments. No other machine we are aware of can do this, certainly none in Merseyside.
*Who will it serve?*
The machine will be part of a facility within the Faculty of Health and Life Sciences, University of Liverpool, and we will promote its use across our own bioscience departments; genetics, tissue engineering, physiology, pharmacology etc and local collaborators in Merseyside research institutes. Initially the applicants will optimise methods and usage, but then our long-term vision is that it will be the first of several parallel recording physiological apparatus and that will be managed under the wing of the University of Liverpool Shared Research Facilities (Liv-SRF) and available to all (following training) alongside other such facilities such as GeneMill, Centre for Cellular Imaging, NMR or MS metabolomics, proteomics and sequencing.
*What are we requesting?*
Much of modern biology is based around identifying large numbers of biomolecules in cells and tissues and the pathways that they influence. Due to extraordinary breakthroughs over recent years this can now be done at astonishing speeds unthinkable in the past. For example single cell RNA-sequencing can now identify almost the entire mRNA complement (~active genes) from 50,000 individual cells in one day (may take a couple of days to analyse the data!). However, one area of biology that is still lagging far behind is physiological recording; recording what changes occur in cell electrical behaviour, movement, survival, contraction etc. These are generally performed by highly skilled experts, but typically at the rate of about one cell per day; once trained and all set up. What we are requesting is a new generation of physiological recording apparatus that can identify physiological changes in hundreds of cells per day. Not up to the speed yet of single cell RNA-sequencing, but a massive advance on what was previously possible for us, or any other labs in the North West of England.
*What can this equipment do that existing equipment cannot?*
Whilst this apparatus would be useful across a range of disciplines from genetics, cell biology neuroscience, ageing, stem cells etc, there are three specific facets we need to highlight.
(1) Existing equipment records only one cell at a time or one recording from many cells all aggregated together. That is painfully slow. This new machine allows for the recording of dozens of cells simultaneously making high throughput electrical and contractility assays possible.
(2) As an extension of (1) this machine will allow one to record individual cells not just faster, but in parallel in a structured fashion so that one can track network activity in real time. Cells within tissues do not behave as individual mavericks doing their own thing; they interact in networks and this machine allows these networks to be analysed in Liverpool for the first time.
(3) This equipment allows analysis of cell integrity, movement and electrical properties all at the same time across dozens of cells in the network and in several parallel experiments. No other machine we are aware of can do this, certainly none in Merseyside.
*Who will it serve?*
The machine will be part of a facility within the Faculty of Health and Life Sciences, University of Liverpool, and we will promote its use across our own bioscience departments; genetics, tissue engineering, physiology, pharmacology etc and local collaborators in Merseyside research institutes. Initially the applicants will optimise methods and usage, but then our long-term vision is that it will be the first of several parallel recording physiological apparatus and that will be managed under the wing of the University of Liverpool Shared Research Facilities (Liv-SRF) and available to all (following training) alongside other such facilities such as GeneMill, Centre for Cellular Imaging, NMR or MS metabolomics, proteomics and sequencing.
Technical Summary
University of Liverpool, with support from the BBSRC has invested in several shared facilities to enhance frontiers bioscience; we have a state-of-the art imaging suite, NMR and mass-spec metabolomic units, proteomics, sequencing services and Gene-Mill. We now seek to purchase a Maestro Pro multi-well multifunctional MEA unit to allow automatic and parallel recording of voltage, contractility, cell viability, migration and proliferation in real time. This apparatus would be added to our existing electrophysiology facility and allow a step change in the depth of mechanistic experiments that would be possible in Liverpool. We will initially develop our usage model with exemplar projects supervised by the applicants from prokaryote to mammalian cells. This will be extended out via workshops and presentations with a well-established shared usage model. Sustainability is ensured by management cross-Faculty, rather than one research group. Initial projects will showcase a wide range of Maestro features across several BBSRC Strategic Priority areas.
I) The platform will allow Users to analyse migration, proliferation and cytotoxicity label-free, in real-time and in multiwell format. This will be more accurate, fast and include temporal information missing from current methods. The device is not just an MEA, but a multi-MEA++.
II) The platform will allow the recording of voltage (eg action potentials) and contractility (if applicable) from monolayers and tissue slices from 100s of electrodes in multiple well dishes. This will be 100s of times faster than considering cells in isolation, but also allow users to capture network activity, conduction velocities and synchronisation all impossible before.
III) The system includes a regulated environmental chamber where temperature and CO2 are monitored, logged and remotely readable.
IV) The platform is designed as a multiuser platform with snap-in barcoded plates so multiple Users can swap experiments in and out.
I) The platform will allow Users to analyse migration, proliferation and cytotoxicity label-free, in real-time and in multiwell format. This will be more accurate, fast and include temporal information missing from current methods. The device is not just an MEA, but a multi-MEA++.
II) The platform will allow the recording of voltage (eg action potentials) and contractility (if applicable) from monolayers and tissue slices from 100s of electrodes in multiple well dishes. This will be 100s of times faster than considering cells in isolation, but also allow users to capture network activity, conduction velocities and synchronisation all impossible before.
III) The system includes a regulated environmental chamber where temperature and CO2 are monitored, logged and remotely readable.
IV) The platform is designed as a multiuser platform with snap-in barcoded plates so multiple Users can swap experiments in and out.
