We're hiring! PhD fellowship on dynamic control in hybrid plasmonic nanopores: road to next generation multiplexed single molecule detection
Within the Marie Skłodowska-Curie Doctoral Networks DYNAMO merging of 6 world-leading academic groups and 1 high tech company, we are offering one full PhD fellowship on plasmonic trapping and enhanced UV label-free single protein detection with plasmonic nanopores.
See the complete details in the description enclosed below. The application website is here.
You may also find it on Academic Positions
• Regular submission deadline: September 6, 2022
• Regular registration deadline: October 4, 2022
• Conference: Oct 25-27, 2022
Effective summary: seeing the intrinsic emission from a single natural protein with ultraviolet optical antennas
One of the ultimate goals of molecular biology is to watch how single proteins work in their native state. The current mainstream approach of single molecule fluorescence relies on introducing external fluorescent markers which can lead to severe issues affecting the experimental results. As an alternative to fluorescence labelling, working in the ultraviolet is appealing to take advantage of the intrinsic autofluorescence naturally present in the vast majority of proteins. However, proteins are orders of magnitude dimmer as compared to conventional fluorescent dyes, so that single protein UV detection has remained a challenge so far. New nanotechnology tools need to be introduced to meet this challenge.
In a recent publication in Nature Communications Ultraviolet optical horn antennas for label-free detection of single proteins, our team introduces a novel optical horn antenna platform for label-free detection of single proteins in the UV with unprecedented resolutions and sensitivity. The approach combines (i) a conical horn reflector for fluorescence collection at ultrahigh angles with (ii) a metal nanoaperture for fluorescence enhancement and background screening. Real-time detection of UV autofluorescence from immobilized and diffusing single label-free proteins is demonstrated, together with experiments monitoring unfolding and dissociation upon denaturation of a widely used protein with single molecule resolution.
Optical horn antennas open up a unique new form of spectroscopy enabling the investigation of single proteins in their native state and in real time. This work provides a leap towards the design of biochemical assays with label-free single protein resolution as well as bright optical nanosources.
Improving single molecule fluorescence detection with metal zero-mode waveguide nanoapertures: recent developements
Jerome will give an invited talk at the Biophysical Society Meeting next week. For those unable to join or for those just curious about it, here is a copy of the presentation slides about using metal nanoapertures to improve single molecule fluorescence detection.
Proteins can feature ultrafast structural dynamics in the nanosecond timescale which are very challenging to measure using conventional techniques such as single-molecule Förster resonance energy transfer (smFRET). So far, the limited fluorescence brightness and the relatively long fluorescence lifetimes have limited the temporal resolution of smFRET to the 10 ns range. Moreover, these ultrafast smFRET measurements require very long acquisition times, often above several hours.
In a recent JACS publication in collaboration with the teams of Ben Shuler and Robert Best, the nanophotonic fluorescence enhancement in zero-mode waveguides is used to push forward the smFRET measurements of ultrafast protein dynamics in the low nanosecond range. The previously inaccessible dynamics of a short intrinsically disordered peptide were probed with nanosecond FCS and smFRET, paving the way to the investigation of very rapid biomolecular dynamics.
- The acquisition time needed for nanosecond FCS is reduced by more than one order of magnitude. Measurements that were taking several hours can now be performed in 20 minutes.
- The experimental data can be compared with all-atom molecular dynamics simulations, and used to further improve the accuracy of the numerical model.
- ZMWs from our group can be easily implemented in other labs with consistent optical performance.