During the rotation the student will learn the main principles correlative microscopy and spectroscopy. The student will learn to obtain vesicles, freeze in vitreous ice and image in cryo-light and electron microscopies. Further cryo-Raman spectroscopy will be used to obtain relevant spectra.
The rotation includes image and spectroscopy analysis as well as data dissemination and statistical skills.
The preliminary work is to obtain the best practice for correlative transfer between systems.
Figure: Pilot study (with University of Bristol). HeLa cell is plunge frozen andimaged A) Optically for 10nm gold/488/EGF or 5nm gold/635/Tfn at -196C or B)Low Magnification Cryo-TEM or C) High magnification cryo-TEM
Dr Cathy Merry, School of Medicine
Dr Victoria James, School of Vetrinary Medicine and Science
Proteoglycans have a protein core with large polysaccharide chains attached. The structure of these chains has immense impact on the function and larger organisation, yet the relationship between structure and function remains mostly illusive.
We hypothesise that vesicle trafficking within the cell determines the external structure of polysaccharides. Understanding the role of vesicle transport and proteoglycan expression, is vital to determining the biological function and relevance of vesicles within the complex tissue environment, particularly their targeted uptake by recipient cells as a means of cell to cell communication.
Here we aim to determine the structure of polysaccharide chains on vesicles isolated from in vitro model systems (vesicles with attached proteoglycans), with the intent of later translation into a cellular diagnostic technique (e.g. on migratory cancers).
Triangle project with:
Victoria James: Do the biophysical properties of exosomes relate to their role as in vivo extracellular messengers?
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