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
Batchelor DVB
(2020)
Nested Nanobubbles for Ultrasound-Triggered Drug Release.
in ACS applied materials & interfaces
Bonnefoy S
(2018)
Biallelic Mutations in LRRC56, Encoding a Protein Associated with Intraflagellar Transport, Cause Mucociliary Clearance and Laterality Defects.
in American journal of human genetics
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
De Pablo J.G.
(2018)
On-chip single cell drug uptake tracking using microfluidic traps and Raman microspectroscopy
in 22nd International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2018
Gala De Pablo J
(2018)
Tandem fluorescence and Raman (fluoRaman) characterisation of a novel photosensitiser in colorectal cancer cell line SW480.
in The Analyst
Gala De Pablo J
(2018)
Biochemical fingerprint of colorectal cancer cell lines using label-free live single-cell Raman spectroscopy.
in Journal of Raman spectroscopy : JRS
Gala De Pablo J
(2020)
Detection and time-tracking activation of a photosensitiser on live single colorectal cancer cells using Raman spectroscopy.
in The Analyst
Hartill VL
(2018)
DNAAF1 links heart laterality with the AAA+ ATPase RUVBL1 and ciliary intraflagellar transport.
in Human molecular genetics
Hollstein R
(2015)
HACE1 deficiency causes an autosomal recessive neurodevelopmental syndrome.
in Journal of medical genetics
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 µm) 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/ |