Analysis of Egg White Protein Composition with Double Nanohole Optical Tweezers.

We use a double nanohole optical tweezer to analyze the protein composition of egg white through analysis of many individual protein trapping events. The proteins are grouped by mass based on two metrics: standard deviation of the trapping laser intensity fluctuations from the protein diffusion and the time constant of these fluctuations coming from the autocorrelation. Quantitative analysis is demonstrated for artificial samples, and then, the approach is applied to real egg white. The composition found from real egg white corresponds well to past reports using gel electrophoresis. This approach differs from past works by allowing for individual protein analysis in heterogeneous solutions without the need for denaturing, labeling, or tethering.

Note: A simple image processing based fiducial auto-alignment method for sample registration.

A simple method for the location and auto-alignment of sample fiducials for sample registration using widely available MATLAB/LabVIEW software is demonstrated. The method is robust, easily implemented, and applicable to a wide variety of experiment types for improved reproducibility and increased setup speed. The software uses image processing to locate and measure the diameter and center point of circular fiducials for distance self-calibration and iterative alignment and can be used with most imaging systems. The method is demonstrated to be fast and reliable in locating and aligning sample fiducials, provided here by a nanofabricated array, with accuracy within the optical resolution of the imaging system. The software was further demonstrated to register, load, and sample the dynamically wetted array.