Geometrically hindered motions of a single large solute (particle or polymer) can be imaged in real time via optical microscopy. The dynamics of fluorescent colloidal particles near surfaces and in porous membranes were monitored using confocal microscopy. A method of analysis to estimate diffusivity of particles in the axial direction by observing their intensity fluctuations was developed. The intensity fluctuations correspond to the Brownian motion of the particles in the axial direction. The method was successful in capturing the hindered diffusion of particles close to surfaces and in pores. This study provides a novel route to monitor the dynamics of particles, including biomacromolecules, near surfaces, through porous substrates and biological tissues. Ionic liquid (IL) as a medium for room temperature preservation of biomacromolecules has been proposed and, to investigate the possibility, physicochemical and enzymatic properties of proteins in the neat hydrophilic IL, ethylmethyl imidazolium ethyl sulfate [EMIM][EtSO4] were studied. Spectroscopic techniques were employed to probe the secondary and tertiary structure of proteins whereas light scattering and viscometry were used to estimate the hydrodynamic size. The secondary structure of the protein was retained in the ionic liquid but the tertiary structure was found to change. Alterations in protein conformation/activity were investigated after transfer of the dissolved protein from the IL to buffer. Further, suitability of ionic liquid gels as protein encapsulation and preservation media was assessed.
| Identifer | oai:union.ndltd.org:UMASS/oai:scholarworks.umass.edu:open_access_dissertations-1266 |
| Date | 01 September 2010 |
| Creators | Bihari, Malvika |
| Publisher | ScholarWorks@UMass Amherst |
| Source Sets | University of Massachusetts, Amherst |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | Open Access Dissertations |
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