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Robustly passivated, gold nanoaperture arrays for single-molecule fluorescence microscopy
Colin D Kinz-Thompson1, Matteo Palma, Dileep K Pulukkunat
1Department of Chemistry, Columbia University , New York, New York 10027, United States.
ACS Nano
|August 31, 2013
Summary
Researchers developed a new surface chemistry for nanoapertures to reduce unwanted biomolecule sticking. This allows for clearer single-molecule fluorescence measurements in complex biological samples.
Area of Science:
- Biophysics
- Nanotechnology
- Surface Chemistry
Background:
- Metal-based nanoapertures enhance optical confinement for single-molecule fluorescence measurements.
- Nonspecific adsorption of biomolecules to nanoaperture surfaces limits sensitivity at physiological concentrations.
Purpose of the Study:
- To develop a selective functionalization strategy for gold nanoaperture arrays.
- To overcome the limitation of nonspecific biomolecule adsorption in single-molecule fluorescence assays.
Main Methods:
- Passivation of metallic cladding with methoxy-terminated, thiol-derivatized polyethylene glycol (PEG).
- Functionalization of silica bottoms with a binary mixture of methoxy- and biotin-terminated, silane-derivatized PEG.
- Utilizing biotin-streptavidin interactions for selective biomolecule tethering.
Main Results:
- Achieved selective tethering of biotinylated target biomolecules to nanoaperture bottoms.
- Significantly reduced nonspecific adsorption of fluorophore-labeled ligand biomolecules.
- Enabled observation of ligand-target binding at background concentrations exceeding 1 μM.
Conclusions:
- The developed selective functionalization chemistry enhances the performance of nanoaperture-based single-molecule fluorescence measurements.
- This approach makes previously challenging biological systems accessible for detailed single-molecule investigations.
- Improved assay sensitivity and specificity are crucial for advancing biomolecular interaction studies.
