Optical Science and Engineering ETDs

Publication Date

Fall 12-16-2023

Abstract

ABSTRACT

The dissertation explores innovative techniques in light sheet microscopy, a pivotal tool in biomedical imaging, to enhance its speed, resolution, and efficiency in capturing dynamic biological processes. Light sheet microscopy allows for quick 3D imaging of biological specimens ranging from cells to organs with high spatiotemporal resolution, large field-of-view, and minimal damage, making it vital for in vivo imaging.

The first project introduces a novel optical concept designed to optimize Axially Swept Light Sheet Microscopy (ASLM). This technique is crucial for imaging specimens ranging from live cells to chemically cleared organs due to its versatility across different immersion media. The project presents an innovative approach that enhances the performance of ASLM-based microscopes by scanning two staggered light sheets while synchronizing the rolling shutter of a scientific camera. This method allows ASLMs to image twice as fast without compromising the detection signal, providing a gentler illumination scheme and improving the imaging speed and detection signal of ASLM-based microscopes. The demonstrated technique is validated through imaging experiments on fluorescent beads and a chemically cleared mouse brain, showcasing its potential in advancing the field of light sheet microscopy.

The second project focuses on axial de-scanning using remote focusing in the detection arm of light-sheet microscopy. This technique is essential for high-speed, high-resolution volumetric imaging without disturbing the biological sample. The project introduces a unique optical design that can descan the axial focus movement in the detection arm of a microscope by overcoming the challenges associated with the polarization of emitted fluorescence. This technique allows for aberration-free, multi-color, volumetric imaging without compromising the fluorescent signal. It is demonstrated by acquiring fast dual-color 4D image stacks, highlighting its potential applications in various microscopy techniques that require adjustable Z-stages for volumetric imaging, such as confocal, 2-photon, and other light sheet variants.

Degree Name

Optical Science and Engineering

Level of Degree

Doctoral

Department Name

Optical Science and Engineering

First Committee Member (Chair)

Tonmoy Chakraborty

Second Committee Member

Keith Lidke

Third Committee Member

Diane S Lidke

Fourth Committee Member

Sheng Liu

Keywords

Fluorescence Microscopy, Remote Focusing, Axially Swept Light Sheet Microscopy (ASLM)

Document Type

Dissertation

Language

English

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