Label‐Free Fluctuation Spectroscopy Based on Coherent Anti‐Stokes Raman Scattering from Bulk Water Molecules

Nanoparticles (NPs) and molecules can be analyzed by inverse fluorescence correlation spectroscopy (iFCS) as they pass through an open detection volume, displacing fractions of the fluorescence‐emitting solution in which they are dissolved. iFCS does not require the NPs or molecules to be labeled. However, fluorophores in μm –mm concentrations are needed for the solution signal. Here, we instead use coherent anti‐Stokes Raman scattering (CARS) from plain water molecules as the signal from the solution. By this fully label‐free approach, termed inverse CARS‐based correlation spectroscopy (iCARS‐CS), NPs that are a few tenths of nm in diameter and at pM concentrations can be analyzed, and their absolute volumes/concentrations can be determined. Likewise, lipid vesicles can be analyzed as they diffuse/flow through the detection volume by using CARS fluctuations from the surrounding water molecules. iCARS–CS could likely offer a broadly applicable, label‐free characterization technique of, for example, NPs, small lipid exosomes, or microparticles in biomolecular diagnostics and screening, and can also utilize CARS signals from biologically relevant media other than water.     

Coherent Anti‐Stokes Emission from Gold Nanorods and its Potential for Imaging Applications

We used coherent anti‐Stokes scattering (CAS) to characterize individual gold nanorods (GNRs) and GNR aggregates. By creating samples with different densities of GNRs on silicon wafer substrates, we were able to determine surface coverage by scanning electron microscopy (SEM) and then correlate the coverage to the CAS intensities of the samples. The observed CAS signal intensity was quadratically dependent on the number of particles. We also examined the CAS signal as a function of the excitation polarization and found that the strongest signals in regularly oriented GNRs were observed when the beam polarization was aligned with the longitudinal axis of the GNRs. Irregularly oriented GNRs exhibited a different scattering pattern to that observed for regularly oriented GNRs. The polarization‐dependent scattering from oriented GNRs showed cos⁶ (θ) behavior. By imaging nanoscale‐sized GNR patterns using CAS and evaluating the results with SEM, we show that CAS can be used for efficient, label‐free imaging of nanoscale metallic particles.