Optical microscopy and scanning probe microscopy techniques have been utilised to acquire high resolution topography and fluorescence images of several biological samples. Applying these techniques to patterned samples and single molecules allow the optical properties of a sample to be investigated near to and below the diffraction limit, allowing emission properties to be correlated with those of topography. Optically active biological samples outside of their cellular environment are prone to photo-degredation and in measuring them a challenge is to ensure that optical measurements can be made before the onset of damage to the fluorophore.
In this study two forms of fluorescence microscope have been utilised with scanning probe techniques of AFM and SNOM. These techniques have been used alongside microcontact printed arrays of fluorescent proteins and photosynthetic light harvesting complexes to address the accuracy of the printing technique and it's applicablity to biological components for future bionanotechnological applications. Furthermore, the periodicity associated with the arrays has been applied to the techniques to address the relative resolutions of the microscopes as well as the samples being a drive behind implimenting biologically friendly components/techniques to the microscopes (such as liquid cells).
Larger structures from photosynthetic bacteria have also been addressed in this study in the form of chlorosomes which are model structures for light harvesting in low light conditions. Studies on the spectral properties of populations of 3 species have been conducted in this work with fluorescence microscopy and it has been shown that populations show small local variations in fluorescence. Furthermore it has been shown that the developed scanning fluorescence technique can be applied to photo senstitive samples successfully with only a small number of cases where spectral properties were affected by the measurement technique.
Using high resolution microscopy techniques this research shows the surface patterning techniques in conjunction with biological samples to have mixed success depending on the sample. It also shows spectral measurements on newly discovered chlorosomes with little photo degredation. It further shows the role that the microscopy techniques have in analysing biological systems in different configurations on substrates
