In collaboration with a group at the University of Durham we have developed a number of custom-designed systems allowing us to image complex cell signalling events in intact and en face vessel preparations.
A summary of these projects are given below. Please follow the link to the collaboration website to learn more about these projects.
Studying endothelial cells, the innermost layer of blood vessels, under physiological conditions is hindered by their inaccessibility.
We developed a miniature wide-field, GRadient-INdex (GRIN) optical probe designed to fit inside an intact pressurized artery. This permits endothelial cell signaling to be imaged with subcellular resolution in a large number (~200) of naturally connected endothelial cells at various pressures. This allows us to study endothelial pressure-dependent mechano-sensitive signalling in a physiologically-relevant model.
Commercial myography systems are often prohibitively expensive considering their relatively simple function, that is maintaining a dissected artery at a given temperature and pressure, and tracking the diameter over time.
We have designed and implemented a low cost alternative to commercial systems, which we call VasoTracker. We have made the design, set up, source code and example datasets freely available so that any lab can easily perform pressure myography.
Visit our website here
Remote Refocus Epifluorescence Microscope
Studying the rapid communication between endothelial and smooth muscle layers is difficult due to slow axial scan speeds of many microscopes, and aberrations caused by movement of the oil used with high magnification lenses.
To get around these problems, this bespoke epifluorescence microscope setup has been designed to use refocussing to scan very rapidly between two focal planes without the movement of an objective lens. At these speeds, we will be able to capture calcium signalling events in both layers of en face arterial preparations practically simultaneously.