Developing cytometry and inertial microfluidic tools to analyse and separate different cell cycle stages of Leishmania mexicana

Howell, Jessie (2024) Developing cytometry and inertial microfluidic tools to analyse and separate different cell cycle stages of Leishmania mexicana. PhD thesis, University of Glasgow.

Abstract

The leishmaniases are a group of devastating diseases which affect around 350 million people, predominantly in developing countries. These diseases are caused by infection with the protozoan parasite Leishmania, yet despite its prevalence, much of the fundamental biology of these parasites is still unknown. One such area of research is the cell cycle, a process which is essential for the replication and survival of this parasite. This process has shown promise as a target for new therapeutic drugs, however, studying the cell cycle to identify such targets is made difficult by the high heterogeneity of cultures of these cells. Cultures contain cells in all stages of the cell cycle, making it difficult to understand and identify molecules controlling this process. While tools are available for synchronising cells at a single stage in the cell cycle, they are often sub-optimal, requiring lengthy processing times or introducing artefacts into the cells and affecting their molecular composition.

This thesis therefore explores inertial microfluidics as an alternative method for cell cycle synchronisation, relying purely on the morphological and mechanical characteristics of the cells to drive separation. Protocols using imaging flow cytometry were developed to provide high-throughput morphological information about the cells. As well as providing insight into the dynamics of the cell cycle, this enabled a deeper understanding of the behaviour of non-spherical cells within inertial microfluidic devices and subsequently an enrichment of desired cell populations. Finally, novel tools for the portable and high-speed imaging of particles within inertial microfluidic devices were demonstrated. As such, this work contributes both protocols and fundamental understanding to both the fields of Leishmania and inertial microfluidic research in the hope that these will prove beneficial to future work.

• Item Type: Thesis (PhD)
• Qualification Level: Doctoral
• Subjects: Q Science > QH Natural history > QH301 Biology; Q Science > QR Microbiology; T Technology > T Technology (General)
• Colleges/Schools: College of Science and Engineering > School of Engineering
• Funder's Name: Engineering and Physical Sciences Research Council (EPSRC)
• Supervisor's Name: Reboud, Professor Julien, Jimenez, Dr. Melanie and Hammarton, Dr. Tansy
• Date of Award: 2024
• Depositing User: Theses Team
• Unique ID: glathesis:2024-84653
• Copyright: Copyright of this thesis is held by the author.
• Date Deposited: 05 Nov 2024 11:18
• Last Modified: 05 Nov 2024 11:20
• Thesis DOI: 10.5525/gla.thesis.84653
• URI: https://theses.gla.ac.uk/id/eprint/84653

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