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Imaging Phantoms for Calibration and Performance Assessment for Multiscale Microscopy
Simar, Lexi
Simar, Lexi
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Abstract
Non-linear optical microscopy, with its unique advantages over conventional laser scanning confocal microscopy, has been widely adopted to study biological processes at the cellular level. However, like all other high-resolution optical imaging techniques, non-linear microscopy suffers from focusing issues due to optical aberrations in the sample due to refractive index mismatch. Optical aberrations distort the wavefront of the excitation beam, causing the focal spot to be larger than the diffraction limit. Since the fluorescence efficiency scales nonlinearly with the profile of the focusing excitation beam, aberrations further degrade the image brightness and resolution. This dissertation explores the trials of imaging the complex biology of glioblastoma multiforme-39 (GBM-39) in attempts to understand the mechanisms of tunneling nanotube (TNT) development and utility. Due to immense issues with aberrations of intrinsic properties of the GBM-39 cells, difficulties with focus on the sample and failure to achieve good resolution, the project pivoted to a new modality and new imaging phantom. This combination of phantom and imaging modality leads to the exploration of calibration standards and imaging phantoms to benchmark the multimodal adaptive optics multiphoton fluorescence microscope (mAO-MPFM) system. Ultimately, the imaging phantom was found to be a solution for a wide array of aberrations to be used across optical microscopy as a calibration standard in two-dimensional and three-dimensional space.
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Date
2025-01-01
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University of Kansas
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This item contains archived web content.
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Simar_ku_0099D_20227.pdf
Adobe PDF, 6.93 MB
- Embargoed until 2176-05-31
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Keywords
Bioengineering, Optics, Biomedical engineering, adaptive optics, calibration standard instrumentation, measurements, multiphoton, pollen, resolution