The elutriator forms part of our tissue culture facility, which is run by a dedicated and highly experienced staff member. The elutriator is routinely used to isolate monocytes from buffy coat, with the monocytes often differentiated into macrophages. Both cell types have been implicated in plaque formation in heart disease, so form a valuable resource for our scientists' experimental models.
HRI has four ultracentrifuges; two floor-based models capable of spinning a rotor at speeds of up to 70,000 rpm, and two benchtop models capable of 150,000 rpm. The ultracentrifuges are commonly used for density gradient separations, with applications including the isolation of HDL or LDL (the "good" and "bad" cholesterols) from whole blood, and the isolation and purification of recombinant adenoviruses and lentiviruses used in gene expression experiments.
Gas chromatography coupled with mass spectrometry (GC/MS) is a powerful technique for the quantification of organic compounds in a variety of sample media. GC/MS is routinely used for the detection of pharmaceuticals and illicit drugs in plasma or urine, and can also be used to detect low abundance metabolites produced during regular or perturbed metabolism. GC/MS can be used to gain understanding into disease mechanisms at a cellular level, and can also be used in disease diagnosis providing suitable marker compounds are known. GC/MS uses high temperatures to transfer samples into the gas phase, and then uses mass spectrometry to detect the weight/mass of the individual compounds as they move through the detector. This detection method enhances the sensitivity of the instrument as the compounds of interest typically have different weights/masses and can be readily distinguished from one another by this technique.
Stopped flow is a technique for measuring the rates of fast chemical and biochemical reactions, and can detect changes in solutions within a few thousandths of a second. In general, two reagents are very rapidly mixed together using a pneumatic drive, and then suddenly ‘stopped’ in the sample cell. The ‘stopped’ sample cell is monitored with light and a sensitive detector that can accurately measure small changes in light intensity as a function of time (typically over milliseconds – seconds). Stopped flow can be a valuable tool in understanding the mechanisms involved in numerous chemical and biochemical processes, including, but not limited to, enzyme reactions, protein-protein interactions, drug-binding processes, protein structure and protein folding.
Dionex ion-exchange chromatography system
The Dionex system is regularly used at HRI to detect ions (atoms or molecules with a positive or negative charge) in complex biological samples, including blood, urine and organs. Examples of ions measured in projects to date include chloride, bromide, nitrite and thiocyanate; the latter is elevated in people who smoke. The measurement of these ion concentrations allows scientists to quickly determine whether their concentrations correlate with measure such as disease activity, oxidative damage, or plaque formation, and thus whether they might be important in the development or progression of disease.
Zeiss Axio Scan.Z1 slide scanner
Our recently purchased Zeiss Axio Scan.Z1 slide scanner compliments our Histology facility, where tissue samples can be prepared stained and imaged entirely in-house. The Axio Scan.Z1 has is capable of both fluorescence and brightfield imaging, and can process up to 100 slides per batch, with fully automated focusing, acquisition, and high-resolution imaging of large areas. For scientists, the automation results in greatly increased productivity, as much less time is spent sitting at the microscope and more time generating important data. The high sensitivity of the optics and cameras also means there are less “false negative” results. The Axio Scan.Z1 is an excellent tool for generating high quality images, that are essential for disseminating results via conferences and publications.
IVIS Lumina XRMS III in-vivo-imaging system
The moorLD2IR high laser Doppler system is used for the acquisition of high-resolution blood flow imaging. It can be used for both clinical and research applications, and allows deeper penetration than other blood flow imaging methods such as laser speckle imaging. This capability makes the laser Doppler highly suitable for visualising blood flow through small blood vessels that are below the tissue surface. The laser Doppler is an essential tool for the study of neovascularisation, which is critical in cardiovascular repair and recovery.