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Debra J Brayshaw

Publications and source records attributed to Debra J Brayshaw.

2 recordsLinked to original sources

Molecular adsorption: early stage surface exploration.

In this study, the atomic force microscope has been employed in force spectroscopy mode to gain information on the interaction between long mucin molecules and a positively charged surface during the first few seconds of interaction. Recent studies have revealed that negatively charged mucin molecules introduced to a positively charged surface are kinetically trapped and bind very rapidly, assuming non-equilibrium conformations. This systematic study of surface dwell times has revealed that significant differences exist in mucin adsorption during the first three seconds of introduction to the surface and provides direct evidence of molecular rearrangement for several seconds before trapping occurs. Limited interactions were recorded at dwell times of less than one second, with increased molecular rearrangement observed between 1.5 and 2.25 s. Increasing the surface dwell time beyond this critical limit caused rupture of the tip-tethered mucin molecules during the retract cycle of the cantilever. All subsequent recorded events, at increased dwell times up to 3s, revealed events at much reduced distances from the point of contact between the mucin functionalised-cantilever and the positively charged surface.

Adsorption↗

Optimisation of sample preparation methods for air imaging of ocular mucins by AFM.

The addition of cations to the imaging buffer for AFM has been previously shown to improve the binding of biological molecules to mica. Investigations were carried out to find the concentration of NiCl(2) required to immobilize mucin molecules on a freshly cleaved mica surface, for imaging using intermittent contact in air. Drop-deposition of samples prepared in HEPES buffer with 1, 2 and 5mM NiCl(2) revealed the sensitivity of the mucin molecules to salt. Dialysis of the mucin solutions dramatically reduced the amount of salt present and allowed single molecules to be imaged, revealing a variation in thickness along their length. Spray deposition of the same mucin solutions produced single molecules that, although less affected by co-adsorbed salt, showed a degree of self-folding. This shows the sensitive balance between HEPES and NiCl(2) required for successful imaging of the sub-molecular features of individual mucin molecules.

Air↗