Light-sheet microscopy is widely recognised in the field of bioimaging as one of the
main options when it comes to 3D imaging of large cleared samples or live imaging
of organisms. This can be mainly attributed to certain advantages that result from its
unique orthogonal optical geometry, such as low photobleaching and fast acquisition
of 3D volumes. This work introduces two compact light-sheet microscopy systems
that explore the miniaturization of the technique, with the use of small-scale devices
that control the operation of the illumination and imaging path. In the two main
systems presented in this work two different 2D micro-electromechanical systems
(MEMS) mirrors are used for the generation of the scanned light-sheet in one axis, as
well as, for the parallel translation of the light-sheet on the orthogonal axis. In this way
the parallel planes of the sample can be illuminated without the need of a mechanical
stage. This type of optical translation is coupled with the use of a tunable lens in the
imaging path that refocuses to each subsequent illuminated plane. The result of this is
a stage-free microscope with all optical scanning. An exploration to the use of a
secondary tunable lens is also presented in this work as a tool for homogenising the
light-sheet thickness throughout the field of view (FOV) by a post processing method
of image “tiling”. Both the MEMS and the tunable lens are small scale devices that not
only contribute in a microscope with compact footprint but can equally reduce the
overall cost of the device with relatively inexpensive pricing. In the same spirit the
device is built with custom 3D printed holders and off-the-shelf optomechanical parts.
The use of the 3D printer has also been investigated in the field of optical component
design, with a custom-made 3D printed prism that was developed to correct aberrations
and allow imaging of microscope slides at an angle. Different test samples are used to
characterize the optical paths of the systems as well as its overall imaging performance.
The collection of z stacks is accomplished with the use of a custom control software
whereas attention is given to different ways of visualizing the 3D stacks that are
acquired with the microscope.
| Date of Award | 9 Dec 2024 |
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| Original language | English |
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| Awarding Institution | - University Of Strathclyde
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| Sponsors | University of Strathclyde |
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| Supervisor | Ralf Bauer (Supervisor) & Deepak Uttamchandani (Supervisor) |
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