Curd, A. orcid.org/0000-0002-3949-7523, Cleasby, A., Baird, M. et al. (1 more author) (2024) Modelling 3D supramolecular structure from sparse single-molecule localisation microscopy data. Journal of Microscopy, 296 (2). pp. 115-120. ISSN 0022-2720
Abstract
Single-molecule localisation microscopy (SMLM) has the potential to reveal the underlying organisation of specific molecules within supramolecular complexes and their conformations, which is not possible with conventional microscope resolution. However, the detection efficiency for fluorescent molecules in cells can be limited in SMLM, even to below 1% in thick and dense samples. Segmentation of individual complexes can also be challenging. To overcome these problems, we have developed a software package termed PERPL: Pattern Extraction from Relative Positions of Localisations. This software assesses the relative likelihoods of models for underlying patterns behind incomplete SMLM data, based on the relative positions of pairs of localisations. We review its principles and demonstrate its use on the 3D lattice of Z-disk proteins in mammalian cardiomyocytes. We find known and novel features at ~20 nm with localisations of less than 1% of the target proteins, using mEos fluorescent protein constructs.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2023 The Authors. Journal of Microscopy published by John Wiley & Sons Ltd on behalf of Royal Microscopical Society. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
Keywords: | relative positions, single-molecule localisation microscopy, sparse data analysis, structural modelling, superresolution microscopy, symmetry, Z-disk |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) > Cell Biology (Leeds) |
Funding Information: | Funder Grant number MRC (Medical Research Council) MR/K015613/1 BBSRC (Biotechnology & Biological Sciences Research Council) BB/S015787/1 |
Depositing User: | Symplectic Publications |
Date Deposited: | 10 Oct 2023 16:02 |
Last Modified: | 09 Dec 2024 16:08 |
Status: | Published |
Publisher: | Wiley |
Identification Number: | 10.1111/jmi.13236 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:204069 |