Fan, Xin (2019) Novel methods for recording and reconstructing images in digital holographic microscopy. PhD thesis, National University of Ireland Maynooth.
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Abstract
The difficulty in visualizing unstained biological cells using brightfield microscopy
has resulted in the development of several specialized imaging techniques that can
enhance the contrast of subcellular features without the need for labeling. Examples
include phase contrast, differential interference microscopy, dark field microscopy,
and Rheinberg illumination. However, these techniques are qualitative in nature and
do not provide any direct measurement of cellular morphology in terms of thickness
or refractive index. Quantitative phase imaging refers to a set of emerging methods
with the potential to provide quantitative real-time measurement of the phase delay
introduced by the specimen with nanometric accuracy and with the same spatial
resolution afforded by brightfield microscopy. Quantitative phase imaging, therefore,
provides a powerful means to study cellular dynamics. Several methods exist for
implementing quantitative phase imaging, which include coherent approaches based
on interferometry known as digital holographic microscopy.
Digital holographic microscopy is an optic-electronic technique that enables the
numerical reconstruction of the complex wave-field reflected from, or transmitted
through, a target with a single capture. Together with phase unwrapping, this method
permits a height profile, a thickness profile, and/or a refractive index profile, to be
extracted, in addition to the reconstruction of the image intensity. Digital holographic
microscopy is unlike classical imaging systems in that one can obtain the focused
image without situating the camera in the focal plane; indeed, it is possible to recover
the complex wave-field at any distance from the camera plane. Therefore, the focus
distance from the image plane to the camera plane can be estimated automatically by
using a focus metric.
The aim of the work presented in this thesis is to develop novel methods for
digital holographic microscopy in order to improve the quantitative analysis of
cellular morphology and detect the nucleus in vivo, together with a number of
numerical process techniques both in amplitude and phase profile. This thesis
includes a number of separate contributions, some relating to novel optical systems
that can be used to record the holograms, and some relating to method of processing
the recorded holograms in order to generate meaningful images.
A low-cost compact portable module is proposed that can be easily integrated
with a brightfield microscope in order to record quantitative phase images. This is
the first of two contributions on novel methods to optically record digital holograms.
The second optical system that is proposed is a novel optical architecture for off-axis
digital holographic microscopy, which allows for continuous change in magnification
and numerical aperture by simply moving the sample. There are also three separate
contributions that deal with numerical methods for the reconstruction of images
recorded using digital holographic microscopy. The first relates to a thorough
examination of the potential for sparsity metrics to be used for autofocusing in
digital holographic microscopy. The last two contributions both relate to new image
processing techniques for label-free color staining of subcellular features using the
quantitative phase image as input. The first method is based on simulated Rheinberg
illumination, while the second method is purely digital and can be related to the
concept of local spatial frequency in the image. Both are shown to provide high
quality color images of diatom cells.
Item Type: | Thesis (PhD) |
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Keywords: | Novel methods; recording; reconstructing; images; digital holographic microscopy; |
Academic Unit: | Faculty of Science and Engineering > Electronic Engineering |
Item ID: | 11200 |
Depositing User: | IR eTheses |
Date Deposited: | 09 Oct 2019 14:14 |
URI: | https://mural.maynoothuniversity.ie/id/eprint/11200 |
Use Licence: | This item is available under a Creative Commons Attribution Non Commercial Share Alike Licence (CC BY-NC-SA). Details of this licence are available here |
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