Triphenoxazoles: utilising UV-excitable molecular fluorophores with tuneable emission for flow cytometry

Sund, Sareena Devi (2024). Triphenoxazoles: utilising UV-excitable molecular fluorophores with tuneable emission for flow cytometry. University of Birmingham. Ph.D.

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Abstract

Fluorescently labelled antibodies are a fundamental tool used in countless biological applications from monitoring surgical operations to identifying and diagnosing diseases. It was in the 1940s that Coons et al.1, 2 reported the method of labelling antibodies with a fluorescent molecule (fluorescein isocyanate) to identify antigens in mammalian tissue, a procedure commonly used today.
This thesis focuses on the development and use of fluorescently labelled antibodies in flow cytometry. Antibodies are labelled with a novel class of fluorescent dyes known as triphenoxazoles, which are excited from ~260 nm to ~425nm and can be chemically tuned to emit from 395 nm to over 600 nm. Conjugating triphenoxazoles to antibodies with a wide range of conditions and additives have been tested to improve performance and achieve specificity, without compromising on functionality. Antibody conjugation, characterisation and evaluation using both conventional and spectral flow cytometry are presented.
The majority of fluorescent dyes used in flow cytometry excite with the violet or higher wavelength lasers, relatively few UV excitable fluorophores are available, and some suffer from intrinsic limitations due to their tandem dye nature. Furthermore, as flow cytometry advances with the development of lasers that operate at even lower wavelengths, there is a rapidly increasing demand for reagents that can be excited by both deep UV (320 nm) and UV (355 nm) laser excitations.

Type of Work:

Thesis (Doctorates > Ph.D.)

Award Type:

Doctorates > Ph.D.

Supervisor(s):