Vision Beyond Optics: Standardization, Evaluation and Innovation for Fluorescence Microscopy in Life Sciences
Authors
Huisman, MaximiliaanFaculty Advisor
David GrunwaldAcademic Program
Interdisciplinary Graduate ProgramUMass Chan Affiliations
RNA Therapeutics InstituteDocument Type
Doctoral DissertationPublication Date
2019-04-01Keywords
microscopynanoscopy
fluorescence
cryogenic
cryo-fluorescence
cryoFM
FM
3DSPEED
SPEED
standardization
PSF
PSF engineering
achromatic
dichroic
multi-color
super-resolution
imaging
imaging standard
optics
adaptive optics
meta-data
meta
MetaMax
calibration
characterization
back-projection
image reconstruction
pseudo-tomography
model-based reconstruction
chromatic aberrations
cryogenic imaging
4D-PSF
calibration tool
Bioimaging and Biomedical Optics
Biological Engineering
Biomedical Devices and Instrumentation
Biophysics
Computer-Aided Engineering and Design
Electrical and Electronics
Electromagnetics and Photonics
Electro-Mechanical Systems
Engineering Physics
Molecular Biology
Nanoscience and Nanotechnology
Numerical Analysis and Computation
Optics
Other Engineering
Systems and Integrative Engineering
Metadata
Show full item recordAbstract
Fluorescence microscopy is an essential tool in biomedical sciences that allows specific molecules to be visualized in the complex and crowded environment of cells. The continuous introduction of new imaging techniques makes microscopes more powerful and versatile, but there is more than meets the eye. In addition to develop- ing new methods, we can work towards getting the most out of existing data and technologies. By harnessing unused potential, this work aims to increase the richness, reliability, and power of fluorescence microscopy data in three key ways: through standardization, evaluation and innovation. A universal standard makes it easier to assess, compare and analyze imaging data – from the level of a single laboratory to the broader life sciences community. We propose a data-standard for fluorescence microscopy that can increase the confidence in experimental results, facilitate the exchange of data, and maximize compatibility with current and future data analysis techniques. Cutting-edge imaging technologies often rely on sophisticated hardware and multi-layered algorithms for reconstruction and analysis. Consequently, the trustworthiness of new methods can be difficult to assess. To evaluate the reliability and limitations of complex methods, quantitative analyses – such as the one present here for the 3D SPEED method – are paramount. The limited resolution of optical microscopes prevents direct observation of macro- molecules like DNA and RNA. We present a multi-color, achromatic, cryogenic fluorescence microscope that has the potential to produce multi-color images with sub-nanometer precision. This innovation would move fluorescence imaging beyond the limitations of optics and into the world of molecular resolution.DOI
10.13028/zq9e-9v53Permanent Link to this Item
http://hdl.handle.net/20.500.14038/31235Notes
This thesis received the 2019 Dean's Award for Outstanding Thesis.
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Licensed under a Creative Commons licenseDistribution License
http://creativecommons.org/licenses/by-nc/4.0/ae974a485f413a2113503eed53cd6c53
10.13028/zq9e-9v53
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