Differential labelling of human sub-cellular compartments with fluorescent dye esters and expansion microscopy.
Sheard TMD , Shakespeare T, Seehra RS, Spencer ME, Suen KM & Jayasinghe I. - Nanoscale. 2023.
Postdoctoral research associate, University of Sheffield, UK
Super-resolution microscopy and cardiac research
Currently, I am a postdoctoral research associate at the University of Sheffield (2021-present). My PhD in Biophysics and Cardiac Biology at the University of Leeds (2016-2021) was funded by the Medical Research Council (MRC), through the Discovery Medicine North Doctoral Training Partnership (DiMeN DTP). I previously completed an MPhil in Adaptive Organismal Biology at the University of Manchester (2015-2016).
My research harnesses biophysical imaging techniques, including fluorescence super-resolution techniques like expansion microscopy (ExM) and electron microscopy, to quantify nanoscale biomolecular structures inside cells and multicellular organisms. By better understanding cellular structures we are able to decipher the structure-function relationships which are fundamental to the physical principles underlying biological systems.
Outside of the lab, I love hiking in the Peak district, running and going to gigs to enjoy live music.
Sheard TMD , Shakespeare T, Seehra RS, Spencer ME, Suen KM & Jayasinghe I. - Nanoscale. 2023.
Sheard TMD , Hurley ME, Colyer J, White E, Norman R, Pervolaraki E, Narayanasamy KK, Hou Y, Kirton HM, Yang Z, Hunter L, Shim J, Clowsley AH, Smith AJ, Baddeley D, Soeller C, Colman MA, Jayasinghe I. - ACS Nano. 2019.
Sheard TMD , Hurley ME, Smith AJ, Colyer J, White E, and Jayasinghe I. - Philosophical Transactions of the Royal Society B. 2022.
Sheard TMD and Jayasinghe I. - Methods in Cell Biology. 2020.
Sheard TMD , Howlett L, Kirton H, Yang Z, Gurrola G, Steele D, Jayasinghe I, Valdivia H, Colyer J - bioRxiv. 2020.
Sheard TMD , Kharche SR, Pinali C, Shiels HA - Journal of Experimental Biology. 2019.
School of Biosciences, University of Sheffield, United Kingdom
Supervisor: Dr Izzy Jayasinghe
School of Biomedical Sciences, University of Leeds, United Kingdom
Thesis: "Molecular-scale analysis of cardiac calcium nanodomains and remodelling using expansion microscopy."
Supervisor: Dr Izzy Jayasinghe
School of Medical Sciences, University of Manchester, United Kingdom
Thesis: "Electron tomography of avian cardiac calcium release units."
Supervisor: Professor Holly Shiels
Faculty of Biological Sciences, University of Leeds, United Kingdom
Thesis: "Electron microscopy of muscle myofibrils."
Supervisor: Professor John Trinick
12 months during PhD: 2019-2020
Research project trialling novel imaging probes for the cardiac ryanodine receptor, using peptide ligands imperacalcin and DPc10. Pre-print published, access here.
eCommerce project evaluating company website, also involved with project management, business planning, intellectual property and marketing.
1 month during PhD: 2018
Studied remodelling in idiopathic dilated cardiomyopathy (IDCM) biopsies from human patients, using 4x ExM. Many thanks to Dr David Crossman for welcoming me into his lab, and to the MRC DiMeN for the flexible fund grant.
1 month: 2015
Conducted reef surveys to improve ecological understanding of the biodiversity of corals and Caribbean reef fish, tackling invasive Lionfish species, and assessing manatee distribution in relation to seagrass.
5 months: 2014
Validating a non-invasive experimental refinement to measure levels of water-borne cortisol in fishtanks, as a less stressful alternative to improve fish welfare. Many thanks to Dr Lynne Sneddon and the SfE.
2023
[13] Hurley ME, Shah SS, Sheard TMD, Kirton HM, Steele D, Gamper N & Jayasinghe I. "Super-resolution analysis of the origins of the elementary events of ER calcium release in dorsal root ganglion neurons". Cells.
[12] Sheard TMD, Shakespeare T, Seehra RS, Spencer ME, Suen KM & Jayasinghe I. "Differential labelling of human sub-cellular compartments with fluorescent dye esters and expansion microscopy". Nanoscale .
[11] Seehra RS, Warrington SJ, Allouis BHK, Sheard TMD, Spencer ME, Shakespeare T, Cadby A, Bose D, Strutt D & Jayasinghe I. "Geometry-preserving Expansion Microscopy microplates enable high fidelity nanoscale distortion mapping". Cell Reports Physical Sciences .
[10] Suen KM, Sheard TMD, Lin C-C, Milonaitytė D, Jayasinghe I, & Ladbury JE. "Expansion Microscopy Reveals Subdomains in C. Elegans Germ Granules". Life Science Alliance.
[9] Hurley ME, White E, Sheard TMD, Steele D, & Jayasinghe I. "Correlative super-resolution analysis of cardiac calcium sparks and their molecular origins in health and disease". Open Biology.
2022
[8] Andersen CB, Zulueta Díaz Y, Kure JL, Eriksen MH, Lovatt AL, Lagerholm CB, Morales SV, Sehayek S, Sheard TMD, Wiseman PW, & Arnspang EC. "The Angiotensin II treatment induce reorganization and changes in the lateral dynamics of Angiotensin II type 1 receptor in the plasma membrane elucidated by Photoactivated localization microscopy combined with image spatial correlation analysis". Analytical Chemistry.
[7] Sheard TMD, Hurley ME, Smith AJ, Colyer J, White E, & Jayasinghe I. "Three-dimensional visualisation of the cardiac ryanodine receptor clusters and the molecular-scale fraying of dyads". Philosophical Transactions of the Royal Society B.
[6] Holmes M, Hurley ME, Sheard TMD, Benson AP, Jayasinghe I & Colman MA. "Increased SERCA2a sub-cellular heterogeneity in right-ventricular heart failure inhibits excitation-contraction coupling and modulates arrhythmogenic dynamics". Philosophical Transactions of the Royal Society B.
2020
[5] Hurley ME, Sheard TMD, Norman R, Kirton HM, Shah SS, Pervolaraki E, Yang Z, Gamper N, White E, Steele D & Jayasinghe I. "A correlative super-resolution protocol to visualise structural underpinnings of fast second-messenger signalling in primary cell types". Methods.
[4] Sheard TMD, Howlett L, Kirton HM, Yang Z, Gurrola G, Steele D, Jayasinghe I, Valdivia HH, & Colyer J. "Peptide ligands of the cardiac ryanodine receptor as super-resolution imaging probes". bioRxiv.
[3] Sheard TMD & Jayasinghe I. "Enhanced Expansion Microscopy to Measure Nanoscale Structural and Biochemical Remodeling in Single Cells". Methods in Cell Biology.
2019
[2] Sheard TMD, Kharche SR, Pinali C, & Shiels HA. "3D ultrastructural organisation of calcium release units in the avian sarcoplasmic reticulum". Journal of Experimental Biology.
[1] Sheard TMD, Hurley ME, Colyer J, White E, Norman R, Pervolaraki E, Narayanasamy KK, Hou Y, Kirton HM, Yang Z, Hunter L, Shim J, Clowsley AH, Smith AJ, Baddeley D, Soeller C, Colman MA & Jayasinghe I. "Three-dimensional and chemical mapping of intracellular signaling nanodomains in health and disease with enhanced expansion microscopy". ACS Nano.
13 11 2023: New paper in Nanoscale: “Differential labelling of human sub-cellular compartments with fluorescent dye esters and expansion microscopy”
10 03 2023: 2023 Biophysical Society Annual Meeting San Diego
03 10 2022: New paper in PTRSB: “3D visualisation of the cardiac ryanodine receptor clusters and the molecular-scale fraying of dyads”
18 11 2021: Doctorate confirmed: PhD reflections
31 01 2021: New beginnings in Sheffield!
17 07 2020: End of Badrilla MRC innovation placement
29 06 2020: Pre-print online: RyR peptides as super-resolution imaging probes
23 06 2020: ExM book chapter published in Methods in Cell Biology
13 05 2020: Expansion microscopy innovations and applications