Biomolecular Mask Formation Using UV Patterened Sam Films
Final trchnical rept. Feb 96-31 Jan 99
COLORADO UNIV AT BOULDER DEPT OF PHYSICS
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The research focus has been on the challenging issues involved in achieving, two-dimensional recrystallization of protein S-layer monomers in large -several hundred micron single crystal areas. To acquire facility with the recrystallization process we first explored the experimental conditions which influence protein crystallization e.g., protein concentration. temperature, pH, ionic strength concentration of various mono- and divalent cations, etc. We attempted crystallization experiments in aqueous suspension or at the airwater interface before trying to reassemble protein monomers directly on a patterned surface. We were able to obtain large - 1 micro x 2 micro, well-ordered, recrystallized protein patches following dialysis. As with the native protein crystal, the recrystallized S-layers showed a square lattice with lattice parameter of 13 nm. We established a reproducible protocol for preparing a solution of S-layer monomers for a recrystallization experiment. We used both thin film carbon substrates on transmission electron microscope TEM grids as well as bulk silicon substrates for a variety of diagnostic recrystallization experiments. Protocols for these experiments were established and numerous experiments carried out, including grid modification float, touch, and time gluteraldehyde crosslinking of S-layer crystal and floatation of the formvar directly on the recrystallization solution. We attempted to extrapolate from our work with TEM grids to recrystallization experiments directly on Si wafers.
- Physical Chemistry
- Metallurgy and Metallography
- Atomic and Molecular Physics and Spectroscopy