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Sandmeyer, Alice: Compact realizations of optical super-resolution microscopy for the life sciences. 2019
Inhalt
Introduction and motivation
Theoretical background
Basics of fluorescence
Conventional microscopy
Definitions of resolution limits
Resolution determination concepts
Wide-field microscopy
Deconvolution methods
Super-resolution microscopy
Stimulated emission depletion microscopy
Single molecule localization microscopy
Super-resolution optical fluctuation imaging
Structured illumination microscopy
Fundamentals of biotechnology
Central dogma of protein biosynthesis
Organelles of the eukaryotic cell
Live-cell imaging of HIV-1 particle dynamics at the virological synapse with a compact 3D fluorescence microscope
Introduction
Fluorescence wide-field microscope for 3D deconvolution
Construction of the optical setup
Communication flowchart between the devices
Data acquisition and image processing
Imaging HIV-1 particles at the virological synapse
Infection pathway and virological synapse
Labeling and imaging strategies
Visualizing the dynamics of the HIV-1 transfer at the VS
Using 3D-SIM to resolve the transfer of HIV-1
Discussion and conclusion
Cost-efficient solution for SOFI with direct camera comparison
Introduction
Direct comparison of camera sensors on a scientific microscope
Microscope setting and data acquisition
Metric comparison of SOFI reconstructions from the different camera chips
Setting up cost-effective SOFI on a home-built system
Verifying SOFI performance of the CMOS sensor
Biplane SOFI approach
Discussion and conclusion
DMD-based SIM visualizes live cell dynamics and reveals morphology of microgels
Introduction
Analysis of blazed grating effect
Coherent illumination of DMDs
Effect of underlying blazed grating
Simulation of the blazed grating effect
Comparison of simulations and experimental results
Construction of a compact and cost-efficient SIM system
SIM images of biological samples
Resolving subdiffraction sized structures in fixed cells
Live-cell imaging
SIM revealing the morphology of microgels
Imaging conditions and modifications
Visualize internal structure
Discussion and conclusion
Comprehensive conclusion and outlook
Bibliography
Materials and methods
Optical setups and data evaluation
Detailed component lists
SOFI analysis
Blazed grating angle determination software
Sample preparation
Calibration sample
HIV-1 imaging
SOFI samples
DMD-SIM samples
Acronyms
Publications
Acknowledgments
Statutory declaration