Robert Schafer

High-precision sizing of nanoparticles by laser transmission spectroscopy

We describe the implementation of precision laser transmission spectroscopy for sizing and counting nanoparticles in suspension. Our apparatus incorporates a tunable laser and balanced optical system that measures light transmission over a wide (210-2300 nm) wavelength range with high precision and sensitivity. Spectral inversion is employed to determine both the particle size distribution and absolute particle density. In this paper we discuss results for particles with sizes (diameters) in the range from 5 to 3000 nm. For polystyrene particles 404 to 1025 nm in size, uncertainties of ±0.5% in size and ±4% in density were obtained. For polystyrene particles from 46 to 3000 nm in size, the dynamic range of the system spans densities from ~10(3)/ml to ~10(10)/ml (5 × 10(-8) to 0.5 vol. %), implying a sensitivity 5 orders of magnitude higher than dynamic light scattering.

Quantitative and Rapid DNA Detection by Laser Transmission Spectroscopy

Laser transmission spectroscopy (LTS) is a quantitative and rapid in vitro technique for measuring the size, shape, and number of nanoparticles in suspension. Here we report on the application of LTS as a novel detection method for species-specific DNA where the presence of one invasive species was differentiated from a closely related invasive sister species. The method employs carboxylated polystyrene nanoparticles functionalized with short DNA fragments that are complimentary to a specific target DNA sequence. In solution, the DNA strands containing targets bind to the tags resulting in a sizable increase in the nanoparticle diameter, which is rapidly and quantitatively measured using LTS. DNA strands that do not contain the target sequence do not bind and produce no size change of the carboxylated beads. The results show that LTS has the potential to become a quantitative and rapid DNA detection method suitable for many real-world applications.

The role of lateral tension in calcium-induced DPPS vesicle rupture.

We assess the role of lateral tension in rupturing anionic dipalmitoylphosphatidyserine (DPPS), neutral dipalmitoylphosphatidylcholine (DPPC), and mixed DPPS-DPPC vesicles. Binding of Ca(2+) is known to have a significant impact on the effective size of DPPS lipids and little effect on the size of DPPC lipids in bilayer structures. In the present work we utilized laser transmission spectroscopy (LTS) to assess the effect of Ca(2+)-induced stress on the stability of the DPPS and DPPC vesicles. The high sensitivity and resolution of LTS has permitted the determination of the size and shape of liposomes in solution. The results indicate a critical size after which DPPS single shell vesicles are no longer stable. Our measurements indicate Ca(2+) promotes bilayer fusion up to a maximum diameter of ca. 320 nm. These observations are consistent with a straightforward free-energy-based model of vesicle rupture involving lateral tension between lipids regulated by the binding of Ca(2+). Our results support a critical role of lateral interactions within lipid bilayers for controlling such processes as the formation of supported bilayer membranes and pore formation in vesicle fusion. Using this free energy model we are able to infer a lower bound for the area dilation modulus for DPPS (252 pN/nm) and demonstrate a substantial free energy increase associated with vesicle rupture.

In situ ion gun cleaning of surface adsorbates and its effect on electrostatic forces

To obtain precise measurements of the Casimir force, it is crucial to take into account the electrostatic interactions that exist between the two boundaries. Two otherwise grounded conductors will continue to have residual electrostatic effects from patch potentials existing on the surfaces. In this paper, we look at the effect of in situ cleaning of adsorbate patches, and the resultant effect on the net electrostatic potential difference between two surfaces. We find a significant reduction in the residual potential due to in situ Ar+ cleaning for the samples used.