Developing a Facility for Sequential Isolation, Manipulation, Observation & Analysis of Single Cells

Lead Research Organisation: University of Leeds
Department Name: School of Medicine

Abstract

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Technical Summary

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Planned Impact

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Publications

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Abou-Saleh RH (2020) Freeze-Dried Therapeutic Microbubbles: Stability and Gas Exchange. in ACS applied bio materials

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Armistead FJ (2023) QCM-D Investigations on Cholesterol-DNA Tethering of Liposomes to Microbubbles for Therapy. in The journal of physical chemistry. B

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Batchelor DVB (2020) Nested Nanobubbles for Ultrasound-Triggered Drug Release. in ACS applied materials & interfaces

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Batchelor DVB (2022) The Influence of Nanobubble Size and Stability on Ultrasound Enhanced Drug Delivery. in Langmuir : the ACS journal of surfaces and colloids

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Bourn MD (2020) High-throughput microfluidics for evaluating microbubble enhanced delivery of cancer therapeutics in spheroid cultures. in Journal of controlled release : official journal of the Controlled Release Society

 
Title Cells under stress: An inertial-shear microfluidic determination of cell behaviour - dataset 
Description The deformability of a cell is the direct result of a complex interplay between the different constituent elements at the subcellular level, coupling a wide range of mechanical responses at different length-scales. Changes to the structure of these components can also alter cell phenotype, thus the critical importance of cell mechano-response for diagnostic applications. The response to mechanical stress depends strongly on the forces experienced by the cell. Here we use cell deformability in both shear-dominant and inertia-dominant microfluidic flow regimes to probe different aspects of the cell structure. In the inertial regime we follow cellular response from (visco-)elastic through plastic deformation to cell structural failure and show a significant drop in cell viability for shear stresses above > 11.8 kN/m2. Comparatively, a shear-dominant regime requires lower applied stresses to achieve higher cell strains. From this regime, deformation traces as a function of time contain a rich source of information including; maximum strain, elastic modulus and cell relaxation times and thus provide a number of markers for distinguishing cell types and potentially disease progression. These results emphasise the benefit of multiple parameter determination for improving detection and will ultimately lead to improved accuracy for diagnosis. We present results for leukemia cells (HL60) as a model circulatory cell as well as for a colorectal cancer cell line SW480 derived from primary adenocarcinoma (Dukes stage B). SW480 were also treated with the actin disrupting drug Latrunculin A (LatA), to test the sensitivity of flow regimes to the cytoskeleton. We show that the shear regime is more sensitive to cytoskeletal changes, and that large strains in the inertial-regime cannot resolve changes to the actin cytoskeleton. 
Type Of Material Database/Collection of data 
Year Produced 2019 
Provided To Others? Yes  
 
Title Dataset associated with "Nested-Nanobubbles for Ultrasound Triggered Drug Release" 
Description Due to their size (1-10 ┬Ám) microbubble-based drug delivery agents suffer from confinement to the vasculature, limiting tumour penetration and potentially reducing drug efficacy. Nanobubbles (NBs) have emerged as promising candidates for ultrasound triggered drug delivery, due to their small size allowing drug delivery complexes to take advantage of the enhanced permeability and retention effect. In this study we describe a simple method for production of Nested-NBs, by encapsulation of nanobubbles (~ 100 nm) within drug loaded liposomes. This method combines the efficient and well-established drug loading capabilities of liposomes, whilst utilising NBs as an acoustic trigger for drug release. Encapsulation was characterised using Transmission Electron Microscopy with encapsulation efficiency of 22 ± 2 %. Nested-NBs demonstrated echogenicity using diagnostic B-mode imaging and acoustic emissions were monitored during high intensity focused ultrasound (HIFU) in addition to monitoring of model drug release. Results showed that although the encapsulated NBs were destroyed by pulsed HIFU (peak negative pressure 1.54 - 4.83 MPa), signified by loss of echogenicity and detection of inertial cavitation, no model drug release was observed. Changing modality to continuous wave (CW) HIFU produced release across a range of peak negative pressures (2.01 - 3.90 MPa), likely due to a synergistic effect of mechanical and increased thermal stimuli. Due to this, we predict that our NBs contain a mixed population of both gaseous and liquid core particles, which upon CW HIFU undergo rapid phase conversion, triggering liposomal drug release. This hypothesis was investigated using previously described models to predict the existence of droplets and their phase change potential and the ability of this phase change to induce liposomal drug release. 
Type Of Material Database/Collection of data 
Year Produced 2020 
Provided To Others? Yes  
URL https://archive.researchdata.leeds.ac.uk/685/
 
Title Dataset associated with 'Biochemical fingerprint of colorectal cancer cell lines using label-free live single-cell Raman spectroscopy' 
Description Raw Raman spectra of the average of each single cell in different colorectal cencer cell lines, and preprocessed data used for multivariate analysis. 
Type Of Material Database/Collection of data 
Year Produced 2018 
Provided To Others? Yes  
 
Title High-throughput Microfluidics for Evaluating Microbubble Enhanced Delivery of Cancer Therapeutics in Spheroid Cultures - Dataset 
Description This collection of datasets provides the data used to create the graphs throughout the associated manuscript. Data for spheroid trapping efficiency (fig 1b), fluid flow rates (fig 2c) and spheroid viabilities and diameters (figs 3b, 4c, 5b and 6c) have all been provided. 
Type Of Material Database/Collection of data 
Year Produced 2020 
Provided To Others? Yes  
URL http://archive.researchdata.leeds.ac.uk/692/