These laboratories provide state of the art facilities for postgraduate biomedical research
These laboratories provide state of the art facilities for postgraduate biomedical research in the areas of biochemistry, biophysics, flow cytometry, histology, mammalian cell and tissue culture, microbiology, microscopy, molecular biology, tissue processing laboratory and other related subjects.
The areas of biomedical research being undertaken are wide-ranging. They include fundamental and translatable investigations of neurodegenerative diseases, cancer biology, infectious diseases, oral diseases, tissue repair and regeneration.
Tissue Culture Suite (DB338)
Suspension, monolayer and 3D culturing techniques of mammalian and human cells and tissue form a central requirement for much of the research carried out within the biomedical research facility. The tissue culture suite comprises of a central area containing Class II laminar flow hoods for the maintenance of cell lines, incubators, centrifuges, and bench space for non-sterile experimental work. A separate dedicated room for establishing and maintaining primary cultures is also available.
Molecular Biology Suite (DB344, DB337)
Laboratory Manager: Dr William Goodwin
The Molecular Biology suite has two separate rooms dedicated to the Pre and Post PCR preparation of samples to minimise contamination that could result in erroneous results. Instrumentation includes Applied Biosystems 2970, Verti and 9700 PCR machines, two real-time PCR platforms – the Roche Light Cycler and Applied Biosystems 7500, Applied Biosystems 310 and 3500 Capillary electrophoresis platforms and Life Technologies Ion Torrent Sequencing machine. Key applications include the quantification of gene expression and detection of a variety of polymorphisms.
General Biochemical Laboratory (DB312)
The General Biochemical Laboratory has bench space for carrying out a wide range of biochemical applications including SDS Page Gel Electrophoresis, Western Blotting and Immunohistochemical staining. The laboratory also contains highly sensitive filter based microplate readers capable of measuring fluorescence (top and bottom), luminescence and absorbance. In addition to single measurements, scanning, kinetic measurements and temperature control are also possible. The Bio-Plex 200 multiplex system, in conjunction with a Bio-Plex Pro II wash station, also provides a flexible, user accessible, microplate-based multiplex immunoassay system, allowing the simultaneous concentrations of up to 100 different analytes (proteins and peptides or nucleic acids) in approximately 50 µl of serum, CSF or cell lysate samples within 3 hours.
Microscopy (DB317, DB331)
Laboratory Manager: Dr Zsolt Fabian
Either a Zeiss LSM 510 Laser Scanning Confocal Microscope or a Modified Zeiss Cell Observer system featuring the Zeiss definite focus, Colibri LED illumination and AxioVision 4 digital image processing software with Axiovision tracking module is available. Both of these systems allow the capture of high resolution single image and z-section acquisition of 3 fluorochromes simultaneously. The cell observer system also incorporates an incubation chamber enabling live cell imaging over several days and a motorised stage which allows imaging of a sample over a tiled area that extends beyond the field of view.
Flow Cytometry (DB316, DB312)
Laboratory Manager: Dr Geraldine O'Connor
The FACSAria Cell Sorter is able to rapidly and simultaneously measure up to 9 different parameters on thousands of individual fluorescently labelled cells within either live or fixed cell populations. Populations of interest can be accurately sorted for subsequent culture and analysis. In addition, the Guava Easycyte HT Benchtop Flow Cytometer, fitted with a robotic sample tray for handling 96 well plates, provides a compact, user friendly and cost effective technique for high throughput analysis.
Biophysics Laboratory (DB323)
Laboratory manager: Dr Sarah R Dennison
The Biophysics Laboratory investigates the structure, function, and activity of bioactive molecules. The experimental approach of this research laboratory is to investigate peptide / lipid interactions using Langmuir Blodgett troughs. This is a powerful technique, which allows biological membranes to be mimicked by lipid monolayers and is able to provide detailed physiochemical and thermodynamic data on the interaction of peptides and other novel pharmaceutical compounds with biological membranes. The laboratory is equipped with 4 NIMA Technology Langmuir Blodgett troughs (KVS-Nima Biolin Scientific Langmuir troughs : NIMA model 102M and 112D Langmuir Trough, NIMA model 611 LB Deposition trough, NIMA 601BAM Langmuir Film Balance Brewster Angle Microscope trough) and 2 custom made 15 ml PTFE troughs.
Circular Dichroism (CD) is one of the most widely used methods for determining structure and monitoring structural changes of biomolecules in solution. The Biophysics laboratory is equipped with a J-815 Jasco Circular Dichroism Spectrometer, which can be used, for example, to investigate protein/peptide folding, protein/peptide-membrane interactions e.g.in the presence of lipid systems mimetic of bacterial/tumour membranes, carbohydrate conformation and investigations into protein-protein interactions.
Fluorescence spectroscopy is another technique used to explore the interactions between peptides or novel pharmaceutical compounds with membranes. These investigations provide information about vesicle leakage and binding during these membrane interactions. The laboratory is equipped with a user-friendly Jasco FP-6500 spectroflourimeter. The main focus of the work in the Biophysics Laboratory is the mechanism underlying antimicrobial peptide (AMP) membrane interaction. It has been suggested that many AMPs use a pore-type mechanism of membrane interaction and although there have been many predictions; very few researchers have been able to show, experimentally. The Nanion port-a-patch is a technique being developed in the biophysics area to investigate membrane channels. This chip-based automated patch-clamp has been used to investigate the permeation of antibiotics through a planar lipid bilayer. This technique has several advantages over the classical montal-muller membrane system, one of which is the high reproducibility of the system. Reproducibility of vesicle preparation is also essential for Biophysics research. The laboratory is also equipped with a Nanion vesicle PrepPro an automated system for the preparation of solvent-free giant unilamellar vesicles (GUVs).
Microbiology Laboratories (DB302 and DB322)
The Microbiology laboratories investigate how bacteria sense and respond to environmental change, focusing particularly on the structure and function of bacterial signal transduction pathways (mainly two-component signal transduction systems) and regulators of bacterial gene expression. Research includes investigations of the intact sensory proteins of these pathways, which are usually located in the bacterial membrane and are responsible for sensing environmental stimuli and stresses, and also their partner cytosolic regulators that effect appropriate adaptive responses (usually by binding to specific promoters and thereby changing gene expression). The principal pathways under investigation are those found in a range of pathogenic bacteria associated with hospital-acquired infections. The main approaches and tools of the research include: molecular biological methods to produce purified signal transduction proteins (produced through heterologous overexpression of the encoding genes in the bacterium E. coli); biophysical methods to investigate ligand-protein interactions of the signal transduction proteins involved, including the intact membrane sensory proteins; elucidation of the three-dimensional structures with a view to structure-based drug design; and phosphorylation-based activity assays to screen and identify candidate signaling ligands and inhibitors. The research currently receives external research funding from BBSRC and the Diamond Light Source Ltd., Oxfordshire.
Investigation of the membrane sensory protein components of these pathways can be technically-challenging due to their inherent hydrophobicity. Expertise in membrane protein biology are required and these are available in the Microbiology facility for expressing and purifying membrane proteins, including: overexpression of the encoding membrane protein genes in large-scale cultures of Escherichia coli hosts; induction of gene expression using chemical inducers; cell harvesting and isolation of membranes using sucrose density gradient centrifugation and ultracentrifugation; and the purification of recombinant proteins with suitably engineered tags from detergent-solubilised membrane preparations using metal affinity chromatography.
The Microbiology Laboratories also provide University facilities for the isolation, identification and investigation of Class 1 and 2 microorganisms. DB322 houses facilities for the preparation and sterilisation of cultivation media and for microorganism isolation, cultivation and preservation. It is located close to autoclave facilities that receive technical support for large-scale media preparation and disposal.
Yeast Laboratory (DB309)
Laboratory Manager: Dr Clare Lawrence
The yeast lab uses a variety of yeast species to undertake research in a number of areas, such as using yeast as a model organism to help study and understand human diseases. This research encompasses a range of scientific disciplines, including cell and molecular biology, biochemistry and microbiology, and multiple techniques. To undertake this work the yeast lab contains a variety of equipment including shaking incubators, a bead beater, a spectrophotometer, DNA and protein gel apparatus and PCR machines.
Main contact: BiomedResearch@uclan.ac.uk
Dr Sarah Dennison | Biomedical Research Facility Manager
+44 (0)1772 89 4475 | Darwin Building DB328
Dr Julie Burrow | Senior Research Assistant – Project Support
+44 (0)1772 89 2482 | Darwin Building DB328
Carly Morris | Research Technician
+44 (0)1772 89 2884 | Darwin Building DB 329
Darwin Building 3rd floor
Facilities open 24/7 (security approved swipe card access or keypad only; restricted to postgraduate students and staff using the facilities)
Industry support (IGA work)