Surface passivation of zero-mode waveguide nanostructures: benchmarking protocols and fluorescent labels

  title={Surface passivation of zero-mode waveguide nanostructures: benchmarking protocols and fluorescent labels},
  author={Satyajit Patra and Mikhail Baibakov and Jean-Beno{\^i}t Claude and J{\'e}r{\^o}me Wenger},
  journal={Scientific Reports},
Zero mode waveguide (ZMW) nanoapertures efficiently confine the light down to the nanometer scale and overcome the diffraction limit in single molecule fluorescence analysis. However, unwanted adhesion of the fluorescent molecules on the ZMW surface can severely hamper the experiments. Therefore a proper surface passivation is required for ZMWs, but information is currently lacking on both the nature of the adhesion phenomenon and the optimization of the different passivation protocols. Here we… 
Fluorescence Brightness, Photostability, and Energy Transfer Enhancement of Immobilized Single Molecules in Zero-Mode Waveguide Nanoapertures
Zero-mode waveguide (ZMW) nanoapertures are widely used to monitor single molecules beyond the range accessible to normal microscopes. However, several aspects of the ZMW influence on the
Long-Range Single-Molecule Förster Resonance Energy Transfer between Alexa Dyes in Zero-Mode Waveguides
It is shown that the FRET enhancement inside a ZMW does not depend on the set of fluorescent dyes, an important step to establish the relevance of ZMWs to extend the sensitivity and detection range of FRET, while preserving its ability to work on regular fluorescent dye pairs.
Zero-mode waveguides can be made better: fluorescence enhancement with rectangular aluminum nanoapertures from the visible to the deep ultraviolet
Nanoapertures milled in metallic films called zero-mode waveguides (ZMWs) overcome the limitations of classical confocal microscopes by enabling single molecule analysis at micromolar concentrations
Purcell radiative rate enhancement of label-free proteins with ultraviolet aluminum plasmonics
The first complete characterization of the Purcell effect and radiative rate enhancement for the UV intrinsic fluorescence of label-free β-galactosidase and streptavidin proteins in plasmonic aluminum nanoapertures is reported.
Ultraviolet Photostability Improvement for Autofluorescence Correlation Spectroscopy on Label-free Proteins.
Combining enzymatic oxygen scavengers with antioxidants and triplet state quenchers greatly promotes the protein photostability, reduces the photobleaching probability and improves the net autofluorescence detection rate, and opens new perspectives to improve the UV durability of organic devices.
Gold Ion Beam Milled Gold Zero-Mode Waveguides
Zero-mode waveguides (ZMWs) are widely used in single molecule fluorescence microscopy for their enhancement of emitted light and the ability to study samples at physiological concentrations. ZMWs
Recent advances in plasmonic nanocavities for single-molecule spectroscopy
This mini-review reports the most recent results on plasmonic nanocavities applied to enhanced single-molecule detection and spectroscopy.
Label-Free Physical Techniques and Methodologies for Proteins Detection in Microfluidic Biosensor Structures
The aim of the present work is to review the variety of physicallabel-free techniques of protein detection and characterization which are suitable for application in microfluidic structures and analyze the technological and material aspects of label-free biosensors that implement these methods.
Fluorolabeling of the PPTase-Related Chemical Tags: Comparative Study of Different Membrane Receptors and Different Fluorophores in the Labeling Reactions
Evaluation of fluorolabeling performance of the chemical tags target of phosphopantetheinyl transferase enzymes (PPTases) once inserted in the sequence of different proteins of interest shows no significant difference in both internalization and axonal transport of the labeled neurotrophins.


Fluorescence energy transfer enhancement in aluminum nanoapertures
Zero-mode waveguides (ZMWs) are confining light into attoliter volumes, enabling single molecule fluorescence experiments at physiological micromolar concentrations. Among the fluorescence
Improved Glass Surface Passivation for Single-Molecule Nanoarrays.
  • H. Cai, S. Wind
  • Chemistry
    Langmuir : the ACS journal of surfaces and colloids
  • 2016
A more versatile method to improve the PEG passivation and enables direct imaging of ordered arrays of single molecules anchored to lithographically patterned arrays of metallic nanodots patterned by e-beam lithography (EBL).
Dark-Field Illumination on Zero-Mode Waveguide/Microfluidic Hybrid Chip Reveals T4 Replisomal Protein Interactions
Conical lens-based dark-field fluorescence microscopy in combination with a ZMW/microfluidic chip for single-molecule fluorescence imaging is presented and it is demonstrated that compared to epi-illumination, the dark- field configuration displayed diminished background and noise and enhanced signal-to-noise ratios.
Deep UV plasmonic enhancement of single protein autofluorescence in zero-mode waveguides.
This work pushes further the limits of plasmonic-enhanced single molecule detection into the UV range and constitutes a major step forward in the ability to interrogate single proteins in their native state at physiological concentrations.
Robustly passivated, gold nanoaperture arrays for single-molecule fluorescence microscopy.
A selective functionalization chemistry is developed whereby the metallic cladding of gold nanoaperture arrays is passivated with methoxy-terminated, thiol-derivatized polyethylene glycol (PEG), and the silica bottoms of those arrays are functionalized with a binary mixture of meth Oxy- and biotin- terminated, silane-dervatized PEG.
Fluorescence enhancement in an over-etched gold zero-mode waveguide.
Results indicate that combining the undercut and reduction of radius of the ZMW can serve as a simple and effective way to essentially improve the performance of an Au-ZMW for single molecule fluorescence detection.
Zero mode waveguides for single-molecule spectroscopy on lipid membranes.
The results presented here are the first reported for supported lipid bilayers in nanostructured devices, and open the possibility of studying membrane imbedded receptors and proteins at physiological concentrations with single-molecule resolution.
A Comparison of Single-Molecule Emission in Aluminum and Gold Zero-Mode Waveguides.
This work indicates that 200 nm gold ZMWs are better suited for single-molecule fluorescence studies in the red region of the visible spectrum, while aluminum appears more suited for the green region ofThe visible spectrum.
Plasmonic antennas and zero-mode waveguides to enhance single molecule fluorescence detection and fluorescence correlation spectroscopy toward physiological concentrations.
Single molecule spectroscopy techniques greatly benefit from ZMW and plasmonic antennas to enter a new dimension of molecular concentration reaching physiological conditions, and is promising to reveal new insights on biological functions and dynamics.
Temperature Measurement in Plasmonic Nanoapertures Used for Optical Trapping
Plasmonic nanoapertures generate strong field gradients enabling efficient optical trapping of nano-objects. However, because the infrared laser used for trapping is also partly absorbed into the