Nanoscale Imaging and Characterisation of Amyloid-β
This thesis presents a method for reliably and robustly producing samples of amyloid-β (Aβ) by capturing them at various stages of aggregation, as well as the results of subsequent imaging with various atomic force microscopy (AFM) methods, all of which add value to the data gathered by collecting information on the peptide’s nanomechanical, elastic, thermal or spectroscopical properties.
Amyloid-β (Aβ) undergoes a hierarchy of aggregation following a structural transition, making it an ideal subject of study using scanning probe microscopy (SPM), dynamic light scattering (DLS) and other physical techniques. By imaging samples of Aβ with Ultrasonic Force Microscopy, a detailed substructure to the morphology is revealed, which correlates well with the most advanced cryo-EM work. Early stage work in the area of thermal and spectroscopical AFM is also presented, and indicates the promise these techniques may hold for imaging sensitive and complex biological materials. This thesis demonstrates that physical techniques can be highly complementary when studying the aggregation of amyloid peptides, and allow the detection of subtle differences in their aggregation processes.
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3 Alzheimers Disease and the Aggregation of Amyloid β
4 Experimental Methodology
5 Substrate Development of the Imaging of Amyloid Proteins with SPM Methods
6 Scanning Probe Microscopy Methods of Imaging Amyloid Peptides During the Aggregation Process
8 The Application of Biophysical Techniques to the Study of the Inhibition of Aggregation of Aβ Using PINPs Liposomes
9 Conclusion and Future Perspectives
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Ab1:42 aggregation AFM-IR Allsop Alzheimer Alzheimer’s disease amplitude amyloid beta-protein amyloid fibrils amyloid peptides amyloid precursor protein amyloid proteins amyloid-beta Applied Physics atomic force microscopy attachment b-sheet beta beta-amyloid Biochemistry Biological Chemistry biological samples Biophysical buffer cantilever Characterisation cleavage cleaved click chemistry Cu(II deflection deposition detected dilution early aggregates elastic fibres frequency hydrogen peroxide imaging increased incubated infrared inhibitor interactions Journal of Biological Kolosov laser lipid liposomes mapping metal ions method mica mid-IR mode Molecular monitored monomers morphology Nano-IR nanomechanical nanoscale neurons oligomeric oligomers particles peptide photothermal plaques PLL-mica population Presenilin probe protofibrils protofilaments response RI-OR2-TAT PINPs sample surface Scanning Probe Microscopy scanning thermal microscopy seen signal smaller aggregates spectroscopy SThM SThM-IR structure substrate techniques thesis Tinker-Mill tip-surface topography toxic UD-PINPs ultrasonic force microscopy ultrasonic vibration wavelengths