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Detecting conformational changes in switchable heterodimer plasmon rulers through iSCAT fluctuation microscopy.

Aidan Oi1, Koustav Kundu1, Ritesh K Bag1

  • 1Department of Chemistry and The Photonics Center, Boston University, Boston, MA 02215, USA. bmr@bu.edu.

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|April 20, 2026
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Summary

Interferometric scattering microscopy detects conformational changes in DNA tethers using Plasmon Rulers. This technique monitors single biopolymer dynamics by observing nanoparticle movement.

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Area of Science:

  • Nanotechnology
  • Biophysics
  • Molecular Biology

Background:

  • Interferometric scattering (iSCAT) microscopy offers high-speed detection of individual nanoparticles.
  • Plasmon Rulers, composed of DNA-tethered noble metal nanoparticles (NPs), exhibit iSCAT contrast dependent on their structure.
  • Fluctuations in iSCAT contrast can reveal conformational dynamics of tether molecules.

Purpose of the Study:

  • To develop and validate heterodimer Plasmon Rulers for detecting conformational changes in DNA tethers.
  • To demonstrate the use of iSCAT fluctuation microscopy for monitoring biopolymer dynamics at the single-molecule level.

Main Methods:

  • Assembled heterodimer Plasmon Rulers from 40 nm and 20 nm gold nanoparticles (Au NPs) with a switchable DNA tether.
  • Incorporated a molecular beacon hairpin in the DNA tether that opens upon binding of micro-RNA (miRNA) 574-3P.
  • Utilized iSCAT microscopy at a 50 kHz acquisition rate to measure heterodimer contrast before and after miRNA exposure.

Main Results:

  • Binding of target miRNA to the molecular beacon increased the conformational space of the tethered 20 nm NP.
  • This increased conformational variability resulted in a significant rise in the mean absolute deviation (MAD) of the iSCAT contrast.
  • The study successfully detected conformational changes in the DNA tether through iSCAT contrast fluctuations.

Conclusions:

  • iSCAT fluctuation microscopy of heterodimer Plasmon Rulers is a viable method for detecting conformational changes in tether molecules.
  • This approach provides a pathway for monitoring the conformational dynamics of individual biopolymers.
  • The study highlights the potential of Plasmon Rulers in nanoscale biophysical investigations.