We report on the investigation on the resolution of optical sensors exploiting Bloch surface waves sustained by one dimensional photonic crystals. A figure of merit is introduced to quantitatively assess the performance of such sensors and its dependency on the geometry and materials of the photonic crystal. We show that the figure of merit and the resolution can be improved by adopting a full ellipsometric phase-sensitive approach. The theoretical predictions are confirmed by experiments in which, for the first time, such type of sensors are operated in the full ellipsometric scheme.
We report on the design, fabrication, and characterization of optical sensors based on Bloch surface waves propagating at the truncation edge of one-dimensional photonic crystals. The sensors can be simultaneously operated in both a label-free mode, where small refractive index changes at the surface are detected, and a fluorescence mode, where the fluorescence from a novel heptamethyne dye label in the proximity of the surface is collected. The two modes operate in the near-infrared spectral range with the same configuration of the optical reading apparatus. The limit of detection is shown to be smaller than that of equivalent surface plasmon sensors and the fluorescence collection efficiency is such that it can be efficiently analyzed by the same camera sensor used for label-free operation.
Photonic crystal (PC) enhanced fluorescence has been proposed as a novel tool for early disease detection in a liquid biopsy format. However, photobleaching of the emitters has never been deeply investigated, despite its cross section is expected to increase due to the large field intensity enhancement. Herein, we report on a comprehensive theoretical description of the stationary fluorescence emission of molecular emitters bound at the surface of a one dimensional photonic crystal (1DPC) biosensor. The model considers coupling of the emission to the large local density of the states provided by the 1DPC, in particular to the Bloch surface waves, which can be characterized by different polarization states. The rotational diffusion equation in the presence of photobleaching was solved analytically by a Laplace spherical harmonics analytical approach. The results show that photobleaching can severely affect the fluorescence emission in terms of total intensity and polarization composition, suggesting that a careful analysis of fluorescence anisotropy in biosensing experiments with PC should be carried out. We applied the model to some case of study conditions that take place in the experiments and propose a procedure to rule out the contribution of photobleaching. Last but not least, we propose fluorescence recovery after orientational photobleaching as a new tool to study rotational diffusion of emitters, or labelled proteins, bound at a surface.
We report on the detection of an angiogenic molecule Vascular Endothelial Growth Factor (VEGF) in different biological matrices by means of a new integrated biosensing platform exploiting the properties of Bloch surface waves. The new platform takes advantage of a tandem configuration, in which both label-free and enhanced fluorescence detection are implemented. Specifically designed one dimensional photonic crystals were deposited directly on disposable and low cost plastic biochips. A direct sandwich immunoassay was used to detect VEGF in buffer, cell culture supernatant and human plasma at low concentration (ng/mL). The platform enabled the detection of VEGF in all three matrices with high resolution, fast turnaround time (30 min) and in close agreement with the results of enzyme linked immunosorbent assays
We report on the use of one-dimensional photonic crystals to detect clinically relevant concentrations of ERBB2/neu/Her2 in cell lysates. ERBB2 is a pivotal breast cancer biomarker and targetable oncogenic driver associated with aggressive breast cancer subtypes. To quantitate soluble ERBB2, we developed an optical platform that combines label-free and fluorescence detection modes. Such platform makes use of a sandwich assay in which the one-dimensional photonic crystals sustaining Bloch surface waves are tailored with a monoclonal antibody for highly specific biological recognition (BSW biochip). In a second step, a second antibody to ERBB2 quantitatively detects the bound analyte. The strategy of the present approach takes advantage of the combination of label-free and fluorescence techniques, making bio-recognition more robust and sensitive. In the fluorescence operation mode, the platform can attain the limit of detection 0.3ng/mL (1.5pM) for ERBB2 in cell lysates. Such resolution meets the international guidelines and recommendations (15ng/mL) for diagnostic ERBB2 assays that in the future may help to more precisely assign therapies counteracting cancer cell proliferation and metastatic spread.
We report on the fabrication and physical characterization of optical biosensors implementing simultaneous label-free and fluorescence detection and taking advantage of the excitation of Bloch surface waves at a photonic crystal's truncation interface. Two types of purposely designed one-dimensional photonic crystals on molded organic substrates with micro-optics were fabricated. These crystals feature either high or low finesse of the Bloch surface wave resonances and were tested on the same optical readout system. The experimental results show that designing biochips with a large resonance quality factor does not necessarily lead in the real case to an improvement of the biosensor performance. The conditions for optimal biochip design and operation of the complete bio-sensing platform are established.
We exploit the excitation of electromagnetic surface waves on high-quality dielectric multilayers to measure the very low extinction coefficient of the structures, with a resolution down to 4·10(-7) and in a simple optical configuration. The effect of exposition to a rhodamine 6G solution in water and ethanol is also reported, including dye adsorption in the layers and bleaching upon resonant excitation.
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