- PhD title: 3D in situ structures of Alzheimer’s pathology
What is your research project on and why is it important?
My research project aims to produce 3D structures of Alzheimer’s disease pathology within patient brain samples that are preserved in a physiological hydrated state. Recent advancements in the field have produced atomic-resolution structures of the proteins that cause Alzheimer’s disease, but to achieve this the proteins must be extracted and purified from the brain tissue. My project aims to produce nanometer-resolution structures of these proteins whilst they remain within the brain tissue in their native environments. This is a really exciting project to work on because it can help confirm whether structures produced from extracted proteins really exist in the same conformations within the tissue, and it allows us to get a better understanding of how these disease-causing proteins are arranged and organized within the brain tissue, and what other components of the cellular environment they may interact with. Answering these questions could be insightful for structure-based drug design, and could give us more clues as to how and why these disease causing proteins accumulate and propagate within brains affected by Alzheimer’s disease.
What facilities and specialist equipment do you use to help you carry out your research?
My project uses a technique called cryo-electron microscopy so I use the Bioimaging Facility and the Electron Microscopy facility at the Astbury Biostructure Laboratory extensively. In the EM facility I use the high-pressure freezer to cryo-preserve brain samples. This machine freezes the brain tissue so fast that the water molecules in the tissue cannot form crystalline ice- instead the tissue is preserved in amorphous ice so that an electron beam can pass through without diffracting. I also use the cryo-fluorescence microscope to image the Alzheimer’s disease pathology in my cryogenically preserved brain samples. Once I have a fluorescent map of the pathology within a brain sample, I use a cryo-ultramicrotome to cut thin (~100 nm thick) cryo-sections so that the brain tissue will be thin enough for an electron beam to pass through it to form a crisp image on a detector at the other side. I use the Titan Krios Cryo-electron microscope to take these images using a technique called cryo-electron tomography: the sample is tilted inside the electron microscope and an image is taken at each tilt. Much like other forms of tomography like a CT scan, we can then computationally reconstruct these 2D images taken from different angles to form a 3D volume of Alzheimer’s pathology within cryo-preserved brain tissue.
What do you particularly enjoy about your research?
I love looking at the 3D volumes of brain tissue I collect. The level of detail you can see and all the stuff we have inside our brain cells is absolutely mind blowing. When you look at these nanometer-resolution volumes of brain tissue sometimes you see cellular structures no one has ever observed before which is insanely exciting, and it can be quite a challenge to figure out what things like that are. Other times you see beautiful textbook worthy examples of organelles we know and love like mitochondria or microtubules. The other thing I particularly like about my project is the direct link to disease- I really hope the work we do will one day be useful in the effort to cure Alzheimer’s disease.
Why did you choose to undertake a PhD at the University of Leeds?
The equipment and level of expertise from facility staff to PIs at Leeds is really exceptional. There are only a few places in the UK that have the equipment required for this project and I am very glad that Leeds is one of them. The research environment at Leeds is one of the biggest reasons I chose to do my PhD here. I did my undergrad here and my final year project involved using the electron microscopy facility and the environment created by the facility staff, PhD students, Post-docs and PI superviours was (and is!) so supportive, it really is one of the biggest reasons I wanted to do my PhD here.
Who are your supervisors? How have they helped you with your research so far?
My main supervisor is Dr Rene Frank and my secondary supervisors are Professor Sheena Radford and Dr Eric Hewitt. They are hugely supportive, knowledgeable and enthusiastic, and they always offer good advice. We have wonderful in-depth discussions and debates in my monthly supervisor meetings which have really helped to shape me as a scientist and have given me confidence and the skills to be an independent researcher. They also provide many internal and external opportunities to present my work which is incredibly useful for shaping a meaningful story from all of your data, and for the valuable opportunity to draw on the expertise of others in different but overlapping fields. All three of them have created a collaborative and supportive environment which I believe I have thrived in.
What are your plans after you complete your PhD?
I hope to do a post-doc using the skills and techniques I have learnt throughout my PhD- I really am enjoying this project and research at Leeds so much that I would love to stay and continue it for a bit longer!
Any links we can use for publications and comms about your work