Accurate sizing of nanoparticles using confocal correlation spectroscopy.
The ability to accurately size low concentrations of nanoscale particles in small volumes is useful for a broad range of disciplines. Here, we characterize confocal correlation spectroscopy (CCS), which is capable of measuring the sizes of both fluorescent and nonfluorescent particles, such as quantum dots, gold colloids, latex spheres, and fluorescent beads. We accurately measured particles ranging in diameter from 11 to 300 nm, a size range that had been difficult to probe, owing to a phenomenon coined biased diffusion that causes diffusion times, or particle size, to deviate as a function of laser power. At low powers, artifacts mimicking biased diffusion are caused by saturation of the detector, which is especially problematic when probing highly fluorescent or highly scattering nanoparticles. However, at higher powers (>1 mW), autocorrelation curves in both resonant and nonresonant conditions show a structure indicative of an increased contribution from longer correlation times coupled with a decrease in shorter correlation times. We propose that this change in the autocorrelation curve is due to the partial trapping of the particles as they transit the probe volume. Furthermore, we found only a slight difference in the effect of biased diffusion when comparing resonant and nonresonant conditions. Simulations suggest the depth of trapping potential necessary for biased diffusion is > 1 k(B)T. Overcoming artifacts from detector saturation and biased diffusion, CCS is particularly advantageous due to its ability to size particles in the small volumes characteristic of microfluidic channels and aqueous microdroplets. We believe the method will find increasing use in a wide range of applications in measuring nanoparticles and macromolecular systems.
Kuyper
Christopher L
CL
Department of Chemistry, University of Washington, Seattle, Washington 98195-1700, USA.
Fujimoto
Bryant S
BS
Zhao
Yiqiong
Y
Schiro
Perry G
PG
Chiu
Daniel T
DT
D003198:Computer Simulation; D049329:Nanostructures / Q000737:chemistry* / Q000648:ultrastructure; D010316:Particle Size; D013057:Spectrum Analysis; D013997:Time Factors