Gregory L. Rorrer

Utilizing nature-created photonic crystals, diatoms, we developed immunoassay with efficiently enhanced fluorescence signals. Testing on standard antibody-antigen models achieved unprecedented detection limits of 10⁻¹⁶ M and 10⁻¹⁴ M using fluorescence spectroscopy and fluorescence imaging respectively.

Diatoms are single‐celled microalgae that possess a nanostructured, porous biosilica shell called a frustule. This study characterized the micro‐photoluminescence (μ‐PL) emission of single living cells of the photosynthetic marine diatom Thalassiosira pseudonana in response to UV laser irradiation at 325 nm using a confocal Raman microscope. The photoluminescence (PL) spectrum had two primary peaks, one centered at 500–510 nm, which was attributed to the frustule biosilica, and a second peak at 680 nm, which was attributed to auto‐fluorescence of photosynthetic pigments. The portion of the μ‐PL emission spectrum associated with biosilica frustule in the single living diatom cell was similar to that from single biosilica frustules isolated from these diatom cells. The PL emission by the biosilica frustule in the living cell emerged only after cells were cultivated to silicon depletion. The discovery of the discovery of PL emission by the frustule biosilica within a single living diatom itself, not just its isolated frustule, opens up future possibilities for living biosensor applications, where the interaction of diatom cells with other molecules can be probed by μ‐PL spectroscopy. Copyright © 2016 John Wiley & Sons, Ltd.

We experimentally demonstrate an ultra‐sensitive immunoassay biosensor using diatom biosilica with self‐assembled plasmonic nanoparticles. As the nature‐created photonic crystal structures, diatoms have been adopted to enhance surface plasmon resonances of metal nanoparticles on the surfaces of diatom frustules and to increase the sensitivity of surface‐enhanced Raman scattering (SERS). In this study, a sandwich SERS immunoassay is developed based on the hybrid plasmonic‐biosilica nanostructured materials that are functionalized with goat anti‐mouse IgG. Our experimental results show that diatom frustules improve the detection limit of mouse IgG to 10 pg/mL, which is ˜100× better than conventional colloidal SERS sensors on flat glass. Ultra‐sensitive immunoassay biosensor using diatom biosilica with self‐assembled plasmonic nanoparticles.

We present an innovative surface-enhanced Raman spectroscopy (SERS) sensor based on a biological-plasmonic hybrid nanostructure by self-assembling silver (Ag) nanoparticles into diatom frustules. The photonic-crystal-like diatom frustules provide a spatially confined electric field with enhanced intensity that can form hybrid photonic-plasmonic modes through the optical coupling with Ag nanoparticles. The experimental results demonstrate 4-6× and 9-12× improvement of sensitivities to detect the Raman dye for resonance and nonresonance SERS sensing, respectively. Such low-cost and high-sensitivity SERS sensors have significant potentials for label-free biosensing.

Diatoms are single-celled photosynthetic algae that make silica shells or "frustules" with intricate features patterned at the nano and microscales. In this study, antibody-functionalized diatom biosilica frustules serve as a biosensor platform for selective and label free antibody-antigen immunocomplex formation by enhanced photoluminescence. Biosilica frustules of 10 micron diameter were isolated from cells of the centric marine diatom Cyclotella sp. They were then mounted on glass and covalently functionalized with the model antibody Rabbit Immunoglobulin G (IgG) to yield a uniform nanostructured surface that selectively binds to its complimentary antigen, Goat anti-Rabbit IgG. Diatom frustules possess an intrinsic capacity to emit blue light when excited with a UV laser light source, a property called photoluminescence. Binding the antibody-functionalized diatom frustule with its complimentary antigen selectively enhanced the intrinsic photoluminescence intensity of the diatom frustule by a factor of three, whereas challenging the antibody-functionalized diatom frustule with a non-complimentary antigen, Goat anti-human IgG did not change the intrinsic photoluminescence intensity. The nucleophilic immunocomplex increases the photoluminescence by donating electrons to non-radiative sites on the photoluminescent diatom biosilica, thereby decreasing non-radiative electron decay and increasing radiative emission. The intensified photoluminescence intensity is correlated to the antigen, goat anti-rabbit IgG concentration, with a binding constant of 2.8 ± 0.7x10^-7 M.

Diatoms are single-celled algae that make silica shells called frustules that possess periodic structures ordered at the micro- and nanoscale. Nanostructured titanium dioxide (TiO₂) was deposited onto the frustule biosilica of the diatom Pinnularia sp. Poly-l-lysine (PLL) conformally adsorbed onto surface of the frustule biosilica. The condensation of soluble Ti-BALDH to TiO₂ by PLL-adsorbed diatom biosilica deposited 1.32 ± 0.17 g TiO₂/g SiO₂ onto the frustule. The periodic pore array of the diatom frustule served as a template for the deposition of the TiO₂ nanoparticles, which completely filled the 200-nm frustule pores and also coated the frustule outer surface. Thermal annealing at 680 °C converted the as-deposited TiO₂ to its anatase form with an average nanocrystal size of 19 nm, as verified by x-ray diffraction, electron diffraction, and SEM/TEM. This is the first reported study of directing the peptide-mediated deposition of TiO₂ into a hierarchical nanostructure using a biologically fabricated template.