Spatio-angular Microscopy

Abstract

Photobleaching and phototoxicity pose a problem in live cell imaging. Fluorescence
imaging induces reactive oxygen species in observed organisms
which can alter the behaviour of the sample. Hence, minimising the light
exposure is an important goal.
We augment a widefield epifluorescence microscope with two spatial light
modulators. By controlling the spatial excitation pattern and the angle of
illumination, we can adapt the illumination to the specimen. In many cases,
this technique will create exposures with reduced excitation of the out-of-focus
fluorophores, resulting in better image quality and less phototoxicity.
My custom software is used to obtain an initial image stack of the specimen.
Subsequent image sections are exposed with excitation patterns that
account for the previous image stack. Depending upon the distribution of
fluorophores, this adaptive exposure can considerably reduce photobleaching
and phototoxicity.

Date of Award	2013
Original language	English
Awarding Institution	King's College London
Supervisor	Rainer Heintzmann (Supervisor) & Klaus Suhling (Supervisor)