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We report, for the first time, the use of a surface plasmon resonance (SPR) fiber-optic immunosensor for selective cellular detection through membrane protein targeting. The sensor architecture lies on gold-coated tilted fiber Bragg gratings (Au-coated TFBGs) photoimprinted in the fiber core via a laser technique. TFBGs operate in the near-infrared wavelength range at ∼1550 nm, yielding optical and SPR sensing characteristics that are advantageous for the analyses of cellular bindings and technical compatibility with relatively low-cost telecommunication-grade measurement devices. In this work, we take consider their numerous assets to figure out their ability to selectively detect intact epithelial cells as analytes in cell suspensions in the range of 2-5 × 10(6) cells mL(-1). For this, the probe was first thermally annealed to ensure a strong adhesion of the metallic coating to the fiber surface. Its surface was then functionalized with specific monoclonal antibodies via alkanethiol self-assembled monolayers (SAMs) against extracellular domain of epidermal growth factor receptors (EGFRs) and characterized by peak force tapping atomic force microscopy. A differential diagnosis has been demonstrated between two model systems. The developed immunosensors were able to monitor, in real time, the specific attachment of single intact cells in concentrations from 3 × 10(6) cells mL(-1). Such results confirm that the developed probe fits the lab-on-fiber technology and has the potential to be used as a disposable device for in situ and real-time clinical diagnosis.

We present the development of a platform for label-free biosensing based on overlayered Long Period Gratings (LPGs) working in transition mode. Nano-scale layers of Polystyrene (PS) with different thicknesses were deposited onto the same LPG to test the performances of the device in different working points of its modified sensitivity characteristic. Adsorption dynamic of biotinylated bovine serum albumin (BBSA) onto the PS overlays was on-line monitored as well as a subsequent streptavidin (SA) binding dynamic on the biotinylated sites of the protein ad-layer. Experimental results show that overlayered LPGs are among the most sensitive refractive index transducers to be employed in label-free biochemical detection and that wide margins of further optimization exist.

We report our latest research results concerning the development of a platform for label-free biosensing based on overlayered Long Period Gratings (LPGs) working in transition mode. The main novelty of this work lies in a multilayer design that allows to decouple the problem of an efficient surface functionalization from that of the tuning in transition region of the cladding modes. An innovative solvent/nonsolvent strategy for the dip-coating technique was developed in order to deposit on the LPG multiple layers of transparent polymers. In particular, a primary coating of atactic polystyrene was used as high refractive index layer to tune the working point of the device in the so-called transition region. In this way, state-of-the-art-competitive sensitivity to surrounding medium refractive index changes was achieved. An extremely thin secondary functional layer of poly(methyl methacrylate-co-methacrylic acid) was deposited onto the primary coating by means of an original identification of selective solvents. This approach allowed to obtain desired functional groups (carboxyls) on the surface of the device for a stable covalent attachment of bioreceptors and minimal perturbation of the optical design. Standard 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide / N-hydrosuccinimide (EDC / NHS) coupling chemistry was used to link streptavidin on the surface of the coated LPG. Highly sensitive real-time monitoring of multiple affinity assays between streptavidin and biotinylated bovine serum albumin was performed by following the shift of the LPGs attenuation bands.

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We present real-time monitoring of multiple biomolecular interactions by means of high refractive index (HRI) coated long period grating (LPG) working in transition region. The coating was deposited by dip-coating as a thin layer of few hundreds nanometers of atactic polystyrene (PS). The HRI coating was used to enhance the LPG sensitivity to refractive index changes of the surrounding medium and its thickness was determined in order to tune the coated device at the desired working point. To allow immobilization of specific biomolecules, the PS overlay surface was functionalized through bovine serum albumin (BSA) and glutaraldehyde. In particular, BSA was directly adsorbed on PS through strong hydrophobic interactions, then glutaraldehyde was linked atop exploiting the lysines side chains. This intermediate layer allowed to covalently immobilize human IgG (antibody) on the device surface. To prove the immobilization method, the interaction between IgG and its proper anti-human IgG (antigen) was studied. The four binding interactions were on-line monitored following the shift of the LPG attenuation band. The experiment also suggests a novel and interesting biofunctionalization approach of unreactive synthetic polymers with applications in immunosensing.