The University of Southampton
University of Southampton Institutional Repository

Microfluidic Devices for the Temporal Resolution of T Lymphocyte Competition

Microfluidic Devices for the Temporal Resolution of T Lymphocyte Competition
Microfluidic Devices for the Temporal Resolution of T Lymphocyte Competition
Every cell in the human body is unique. In the immune system this is essential so that a wide range of invading pathogens can be recognised. To achieve the heterogeneity in lymphocytes, immature T lymphocytes and B lymphocytes undergo ‘priming’ in the thymus and bone marrow to diversify the cell surface receptors. As such, bulk analysis of immune cells and overlooking outliers, such as cells exhibiting unusual characteristics or responses, could lead to interesting cell characteristics being disregarded.

By tracking the activation profile of a T lymphocyte against an antigen presenting cell in a high-throughput manner at the single cell level, the variation in activation levels of individual T lymphocytes will be identified. This will lead to an improved understanding of why certain T lymphocytes are more efficient at eradicating diseases.

This thesis describes the design and development of a bespoke microfluidic device able to trap thousands of individual cells in nanolitre wells, for the analysis of cell-cell interactions. The device is optically transparent, enabling T lymphocyte activation to be observed in real time using fluorescent microscopy and calcium staining; thousands of cells may be observed within a single microscope field of view. Cell-cell contact is controllable by the device, allowing the activation of T lymphocytes against antigen presenting cells to be viewed in real time, in a high-throughput manner.

The device has been used to investigate T lymphocyte activation against soluble stimulants, as well as antigen presenting cells. Specifically, the temporal calcium response of single cells has been studied and experimental results are presented. The findings reveal differences in activation profiles of individual cells within a clone population, which are not evident using state-of-the art bulk cell analysis techniques such as flow cytometry. This work highlights the importance of assessing single cell responses in an immune interaction.
University of Southampton
Bateman, Faith
8d8813b8-25ee-4cd3-a84b-2ecfbad1d448
Bateman, Faith
8d8813b8-25ee-4cd3-a84b-2ecfbad1d448
Morgan, Hywel
de00d59f-a5a2-48c4-a99a-1d5dd7854174

Bateman, Faith (2016) Microfluidic Devices for the Temporal Resolution of T Lymphocyte Competition. University of Southampton, Doctoral Thesis, 225pp.

Record type: Thesis (Doctoral)

Abstract

Every cell in the human body is unique. In the immune system this is essential so that a wide range of invading pathogens can be recognised. To achieve the heterogeneity in lymphocytes, immature T lymphocytes and B lymphocytes undergo ‘priming’ in the thymus and bone marrow to diversify the cell surface receptors. As such, bulk analysis of immune cells and overlooking outliers, such as cells exhibiting unusual characteristics or responses, could lead to interesting cell characteristics being disregarded.

By tracking the activation profile of a T lymphocyte against an antigen presenting cell in a high-throughput manner at the single cell level, the variation in activation levels of individual T lymphocytes will be identified. This will lead to an improved understanding of why certain T lymphocytes are more efficient at eradicating diseases.

This thesis describes the design and development of a bespoke microfluidic device able to trap thousands of individual cells in nanolitre wells, for the analysis of cell-cell interactions. The device is optically transparent, enabling T lymphocyte activation to be observed in real time using fluorescent microscopy and calcium staining; thousands of cells may be observed within a single microscope field of view. Cell-cell contact is controllable by the device, allowing the activation of T lymphocytes against antigen presenting cells to be viewed in real time, in a high-throughput manner.

The device has been used to investigate T lymphocyte activation against soluble stimulants, as well as antigen presenting cells. Specifically, the temporal calcium response of single cells has been studied and experimental results are presented. The findings reveal differences in activation profiles of individual cells within a clone population, which are not evident using state-of-the art bulk cell analysis techniques such as flow cytometry. This work highlights the importance of assessing single cell responses in an immune interaction.

Text
Final thesis - Version of Record
Available under License University of Southampton Thesis Licence.
Download (45MB)

More information

Published date: 25 September 2016

Identifiers

Local EPrints ID: 419066
URI: http://eprints.soton.ac.uk/id/eprint/419066
PURE UUID: 71f0fcbb-901b-4467-a506-e6237cebc987
ORCID for Hywel Morgan: ORCID iD orcid.org/0000-0003-4850-5676

Catalogue record

Date deposited: 28 Mar 2018 16:30
Last modified: 14 Mar 2019 05:10

Export record

Download statistics

Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.

View more statistics

Atom RSS 1.0 RSS 2.0

Contact ePrints Soton: eprints@soton.ac.uk

ePrints Soton supports OAI 2.0 with a base URL of http://eprints.soton.ac.uk/cgi/oai2

This repository has been built using EPrints software, developed at the University of Southampton, but available to everyone to use.

We use cookies to ensure that we give you the best experience on our website. If you continue without changing your settings, we will assume that you are happy to receive cookies on the University of Southampton website.

×