Next-Generation, 3-Dimensional Super-Resolution Microscopy

Lead Research Organisation: UNIVERSITY OF CAMBRIDGE
Department Name: Chemistry

Abstract

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Publications

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Studentship Projects

Project Reference Relationship Related To Start End Student Name
EP/N509620/1 30/09/2016 29/09/2022
1943723 Studentship EP/N509620/1 30/09/2017 29/09/2021 Anoushka Handa
 
Description Quantitative imaging in complex biological samples such as brain tissue requires techniques such as super-resolution imaging to better understand the morphology and stoichiometry of proteins, specifically synaptic proteins below the diffraction limit. The 3D double helix-point spread function (DH-PSF) is a technique which has an increased depth of field (~4 µm) and is capable of isotropic resolutions of 25 nm. This makes it highly compatible to investigate sub-synaptic diversity in a physiologically relevant environment. This project uses postsynaptic density 95 (PSD95), a scaffolding protein, genetically expressed to mEos2 which is then imaged in brain tissue using the 3D DH-PSF. PSD95 is known to form nanoclusters which make up the basic structural unit of an excitatory synapse. A deeper understanding of how PSD95 nanoclusters form and how mutations occur in these synapses in the hippocampus contributes knowledge to understanding how this can lead to schizophrenia, learning disabilities and autism. We are halfway through this project which is currently going well however it is too early to discuss results.
Exploitation Route 3D super-resolution imaging provides a deeper understanding into biological samples such as synaptic proteins, . The outcomes of this award will give both biologists and microscopists a better understanding of how clustering of this specific protein (PSD95) forms within the brain as well as the ability to resolve biolgical tissue in 3D down to resolutions of 25 nm. As biological samples in nature are intrinsically 3D, 3D imaging provides a better understanding of many more molecular characteristics of a biological sample in vivo.
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