LSPR Biosensing Approach for the Detection of Microtubule Nucleation.

Microtubules are dynamic protein filaments that are involved in a number of cellular processes. Here, we report the development of a novel localized surface plasmon resonance (LSPR) biosensing approach for investigating one aspect of microtubule dynamics that is not well understood, namely, nucleation. Using a modified Mie theory with radially variable refractive index, we construct a theoretical model to describe the optical response of gold nanoparticles when microtubules form around them. The model predicts that the extinction maximum wavelength is sensitive to a change in the local refractive index induced by microtubule nucleation within a few tens of nanometers from the nanoparticle surface, but insensitive to a change in the refractive index outside this region caused by microtubule elongation. As a proof of concept to demonstrate that LSPR can be used for detecting microtubule nucleation experimentally, we induce spontaneous microtubule formation around gold nanoparticles by immobilizing tubulin subunits on the nanoparticles. We find that, consistent with the theoretical model, there is a redshift in the extinction maximum wavelength upon the formation of short microtubules around the nanoparticles, but no significant change in maximum wavelength when the microtubules are elongated. We also perform kinetic experiments and demonstrate that the maximum wavelength is sensitive to the microtubule nuclei assembly even when microtubules are too small to be detected from an optical density measurement.