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Automated System for Single Molecule Fluorescence Measurements of Surface-immobilized Biomolecules

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Optimal strategy for trapping single fluorescent molecules in solution using the ABEL trap.

Q Wang¹, W E Moerner

¹Departments of Electrical Engineering and Chemistry, Stanford University, Stanford, CA 94305, USA.

Applied Physics. B, Lasers and Optics

|April 13, 2010

Summary

Researchers developed a new Anti-Brownian Electrokinetic (ABEL) trap controller for precisely trapping and analyzing single fluorescent molecules. This innovation improves the ability to study the diffusion of tiny particles in solution.

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Area of Science:

Biophysics
Optical trapping
Nanotechnology

Background:

Trapping small, fluorescent biomolecules in solution is challenging due to their high diffusion coefficients.
Existing methods using the Anti-Brownian Electrokinetic (ABEL) trap require faster, more efficient position sensing for nanometer-sized objects.
Single molecules have limited photon emission, necessitating efficient photon utilization.

Purpose of the Study:

To develop an improved controller for the ABEL trap to enhance the trapping and analysis of single fluorescent biomolecules.
To implement advanced algorithms for faster position sensing and more accurate diffusion coefficient extraction.

Main Methods:

Designed a new ABEL trap controller utilizing a knight's tour scanning pattern for the excitation beam on a 2D lattice.

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Incorporated a Kalman filter-based estimator for optimal real-time position sensing.

Developed a maximum-likelihood-based method for diffusion coefficient extraction.

Main Results:

The new controller design enables tighter trapping of 10-nm-sized single fluorescent biomolecules.
Monte Carlo simulations demonstrate superior performance compared to the simple rotating beam design.
The improved approach significantly enhances the ability to extract diffusion coefficients accurately.

Conclusions:

The novel ABEL trap controller with advanced scanning and filtering significantly improves single-molecule trapping and characterization.
This technology offers a more effective method for studying the dynamics of small particles in solution.
The findings pave the way for more precise investigations in biophysics and nanotechnology.