Mr. John Greenwood
The SITraN Drug Screening Laboratory is associated with Mr John Greenwood who refused to let Motor Neurone Disease destroy his zest for life. Mr Greenwood strongly believed in that science, given time and investment, would deliver a treatment.
As part of its translational research strategy, SITraN has established a Drug Screening Laboratory with state-of-the-art equipment to undertake a comprehensive drug discovery programme for neurodegenerative disorders. Our equipment allows optimized storage of our test drugs and compounds in a fully automated system using dry nitrogen. High-throughput liquid dispensing systems allow large scale screening experiments to be conducted rapidly and with minimal, and thus most efficient, use of the test compounds.
With our automated plate readers, InCell 2000, Opera Phenix and Metabolic Analysis equipment we can further test the effects of the drugs on mitochondrial function, protein distribution, as well as cell size and shape (morphology). The Opera Phenix is our most recent acquisition and this extends the level of detail we can image taking our high content screening potential (previously just the InCell 2000) to new levels.
We screen compound libraries that contain FDA approved drugs and plant extracts for neuro-protective effects and re-purposing of safe drugs. Our hits are also used for proof of concept, validating our targets. We further screen novel compounds and potential therapies developed in our own laboratories.
We collaborate with partners from industry, currently Astra Zeneca, UK, to test novel drugs for therapeutic potential in the treatment of neurodegenerative disorders on a range of established disease models from cell line reporter models (e.g. Nrf2 activation) to human skin cells (fibroblasts) to zebrafish.
The Roylan Multi-pod Storage System:
This system enables our drug libraries to be stored in optimal conditions, avoiding the need for the compounds to be frozen. This minimises the risk of precipitation and degradation through repeated freeze thaw cycles. The system is fully automated and uses dry inert nitrogen gas to remove oxygen and humidity from the system, thus reducing potential damage to the test drugs. Storing the drug libraries in this manner extends the life of the compounds.
The Labcyte Echo 550:
This liquid dispensing system which uses sonic technology to dispense very small volumes (2.5 nanolitres, approximately a millionth of a teaspoon) of compounds from our drug libraries. The system is efficient and economical as it requires no plastic consumables, thus reducing the risk of contamination and loss during screening experiments. The system can perform drug dose response experiments in real time and is used for compound library management as it can measure the volume and quantify the water content of the compounds being tested.
Thermo Scientific Matrix Plate Mate Plus:
This high throughput liquid dispensing system is time efficient, accurate (0.1-300µl) and enables reproducibility and use of standardized operating protocols. This machine supports large scale screening experiments to be conducted with minimal resource use. It complements the acoustic system by being fast and accurate at repetitive dispensing of microliter volumes.
Seahorse Bioscience XF 24 Extracellular Flux Analyser:
This system measures small changes in metabolism using sensitive oxygen and pH electrodes. This system is unique as it is non-invasive and requires less material due to a microenvironment of 7 µl.
The cell samples can then be further tested in other experiments. Alternative methods do not allow metabolic activity and morphology measurements on the same sample.
GE Healthcare InCell Analyser 2000:
This high content screening machine combines the strengths of a automated plate reader with a high power microscope. This enables assay quantification through fluorescence spectroscopy and scrutiny of individual cell morphology. The system measures multiple parameters, providing in depth data on the molecular and metabolic interactions being studied.
PerkinElmer Opera Phenix:
This high content screening machine has all the attributes of the InCell 2000 above, but has the added bonus of confocality and multi cameras. Confocality means the light is focused as well as the objective image allowing detection of smaller biology, essential for identifying some of the most recently identified pathological hallmarks in MND. It also has 2 cameras for simultaneous image detection, decreasing capture time, increasing our through put.
Other Equipment Available:
We also have bulk liquid dispensers for cell plating and back filling plates.
We have a standard fluorescent/luminescent plate reader and a top end plate reader capable of fluorescent polarisation and alpha screening which provides greater sensitivity.
Director of Facility:
Professor Pamela Shaw
Senior Scientific Officer:
Dr Adrian Higginbottom
Contact Adrian Higginbottom for further details:
Telephone: 0114 222 253
Sheffield Institute for Translational Neuroscience
University of Sheffield
385a Glossop Road