Synchronized quartz crystal microbalance and nanoplasmonic sensing of biomolecular recognition reactions.

We present a method providing synchronized measurements using the two techniques: quartz crystal microbalance with dissipation (QCM-D) monitoring and localized surface plasmon resonance (LSPR). This was achieved by letting a thin gold film perforated with short-ranged ordered plasmon-active nanoholes act as one of the electrodes of a QCM-D crystal. This enabled transmission-mode optical spectroscopy to be used to temporally resolve colorimetric changes of the LSPR active substrate induced upon biomolecular binding events. The LSPR response could thus be compared with simultaneously obtained changes in resonance frequency, Deltaf, and energy dissipation, DeltaD, of the QCM-D device. Since the LSPR technique is preferentially sensitive to changes within the voids of the nanoholes, while the QCM-D technique is preferentially sensitive to reactions on the planar region between the holes, a surface chemistry providing the same binding kinetics on both gold and silica was used. This was achieved by coating the substrate with poly(L-lysine)-graft-poly(ethylene glycol) (PLL-g-PEG), which was shown to bind in the same manner on silica and gold modified with a carboxyl-terminated thiol. In this way, the combined setup provided new information about structural changes upon PLL-g-PEG adsorption. We also demonstrate subsequent binding of NeutrAvidin and an immunoreaction utilizing biotin-modified IgG. The combined information from the synchronized measurements was also used in a new way to estimate the sensing volume of the LSPR sensor.