MS Thesis Defense: A. Bajor - 3D Multifocus-Structured Illumination Microscopy. Super resolved 3D fluorescence imaging using diffractive optics
Commercial 3D SIM microscopes are available from several vendors but are expensive and typically only available in academic imaging facilities. Challenges in application of 3D SIM to study living biological specimens include the limited 3D acquisition time required to record each 3D SIM image volume. Multi-focus SIM (MF-SIM) has been previously demonstrated in a proof of concept experiment on a fixed specimen in a single color. My thesis is part of a research project to provide a live specimen compatible MF-SIM system.
My work has included leveraging modern electronic and opto-mechanical devices, alternative illumination and data reconstruction strategies to reduce the complexity of the illumination path, by removing any unnecessary moving or modulated devices, and utilizing a modern phase-based Liquid Crystal on Silicon (LCoS) Structured Light Modulator (SLM) utilizing calibrated sinusoidal phase grating patterns to avoid spurious orders generated from aliased lattice patterns caused by binary gratings.
This approach frees us of some mechanical components and allows us to extend the functionality of our SIM system to easily accept two or more optimized SIM patterns which can be interleaved for multi color acquisitions. This allows us to run samples with dual fluorophores and study the dynamics and interactions of fine structures in samples such as the synaptonemal complex in the model organism \textit{C. elegans.}
Combining 3D SIM with Multifocus Microscopy (MFM) techniques which utilize specialized custom diffractive optics. This approach gives us the ability to simultaneously image 7, 9, or more planes while running a 3D SIM pattern. This greatly speeds up our SIM temporal resolution at the expense of the total intensity being split between the focal planes. Due to the fundamental difference between translating the sample axially through a modulation field, and collecting multiple focal planes sitting in a modulation field, a novel approach is required to recover the encoded axial data and perform a true 3D reconstruction. And is the culmination of this research.
Event Host: Antone Bajor, M.S. Student, Electrical Engineering
Advisor: Sara Abrahamsson
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Wednesday, June 14, 2023 at 4:00pm
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