Next-Generation, 3-Dimensional Super-Resolution Microscopy
Lead Research Organisation:
University of Cambridge
Department Name: Chemistry
Abstract
Super-resolution microscopy is an advanced, interdisciplinary, optical imaging technique currently attracting immense interest as a new and exciting way to break the theoretical diffraction limit of visible light (~250nm). Using single molecule control, super-resolution microscopy retains the non-invasive advantages of fluorescence imaging but has the ability to resolve biological structures more compatible with the spatial scale that these events actually take place on, typically attaining resolutions of ~15nm or better. However what is currently lacking in the field is development of next generation fluorophores that will enable us to address challenges in probing biological systems, primarily due to a lack of understanding about how these molecules function. This project is focusing on fundamental physical chemistry, which underpins this research field. The aim is to focus more on the application to real world biological problems. The aim for this project will be to develop techniques in supramolecular imaging for fluorophores and chromophores under realistic environments to investigate biological systems of real interest, for example to visualise the molecular mechanisms associated with the protein aggregation processes implicated in diseases such as Parkinson's or Alzheimer's.
Organisations
People |
ORCID iD |
| Steven Lee (Primary Supervisor) | |
| Anoushka Handa (Student) |
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| EP/N509620/1 | 01/10/2016 | 30/09/2022 | |||
| 1943723 | Studentship | EP/N509620/1 | 01/10/2017 | 30/09/2021 | 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. |
| Sectors | Other |