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Implementation of a Reference Interferometer for Nanodetection
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A Highly Compensated Interferometer for Biochemical Analysis.

Michael N Kammer1,2, Amanda K Kussrow1, Ian R Olmsted1

  • 1Department of Chemistry and Vanderbilt Institute of Chemical Biology , Vanderbilt University , Nashville , Tennessee 37235 , United States.

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|July 10, 2018
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Summary
This summary is machine-generated.

A novel compensated backscattering interferometer (CBSI) allows sensitive nanovolume refractive index measurements and molecular assays without temperature control. This method achieves high precision even with significant thermal changes, enabling label-free biochemical analysis.

Keywords:
compensated interferometryfree-solution assaylabel-free determinationsmicrofluidicstemperature insensitivity

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Area of Science:

  • * Optical sensing
  • * Biophysical chemistry
  • * Microfluidics

Background:

  • * Accurate refractive index (RI) measurements are crucial for biochemical assays.
  • * Conventional interferometric methods often require precise temperature control, limiting their application.
  • * Nanovolume sample analysis presents challenges in sensitivity and stability.

Purpose of the Study:

  • * To develop an improved interferometric sensing approach for high-sensitivity nanovolume RI measurements.
  • * To enable molecular interaction assays without the need for a temperature controller.
  • * To demonstrate label-free, free-solution biochemical assays at nanomolar concentrations.

Main Methods:

  • * Utilized a compensated backscattering interferometer (CBSI) incorporating a He-Ne laser, microfluidic chip, and CCD array.
  • * Employed a novel method of elongating the laser beam and measuring interference pattern shifts for thermal perturbation compensation.
  • * Separated sample and reference solutions using an air gap or oil droplet for differential RI measurement.

Main Results:

  • * Achieved differential RI measurements with a compensation level of ~5 × 10-8 RIU despite 8 °C thermal fluctuations.
  • * Demonstrated a detection limit of 7 × 10-7 RIU for RI difference measurements.
  • * Successfully quantified binding affinities for mannose-concanavalin A and Ca2+-recoverin interactions at the 10s of nM level.

Conclusions:

  • * The CBSI provides a robust platform for sensitive RI measurements and molecular assays in nanoliter volumes.
  • * The system effectively compensates for thermal perturbations, eliminating the need for stringent temperature control.
  • * CBSI facilitates label-free, free-solution biochemical analysis, opening avenues for various biological studies.