The integration of the Whispering Gallery Modes (WGMs) resonators in a microfluidics platform represents an important feature towards the realization of a compact high performance label-free biosensor. These hollow resonant microstructures present the advantage to combine the WGM resonator properties with the intrinsic capability of integrated microfluidics. In this sense, optical microbubble resonators (OMBRs), intended as a hollow core spherical bulge realized in a glass microcapillary by a suitable fabrication process, with their high Q factors (< 107 in air) well satisfy this requirement. Their operation is based on the fact that, given a small enough wall thickness of the bubble, the WGM optical field extends on both sides of the wall, so that it is possible to couple light into the resonator from an outer waveguide, and at the same time to have interaction of the WGM field with the inner fluid and analyte.
The biosensing mechanism of these devices is based on the WGMs morphological dependence: any change on the OMBR inner surface, due to some chemical and/or biochemical binding, causes a shift of the resonance position and reduces the Q factor of the OMBR. By measuring these changes, important information about the sensing capability of the device can be obtained.
In order to develop an OMBR based biosensor and optimize its performance, a crucial step is represented by its chemical/biochemical functionalization. Here we present a novel technique able to guarantee that the chemical interaction occurs in the OMBR inner wall, leaving the other microfluidic parts completely inert from a biochemical point of view. The method is based on UV photoactivation, which allows to localize the biolayers only in correspondence of the OMBR inner wall. As a proof of concept, an immunoassay based on rabbit IgG/anti rabbit-IgG interaction was performed and. The anti rabbit-IgG antibody was labelled with Alexa Fluor 488 to verify, by a fluorescence characterization, the goodness of this procedure. Moreover, an anti mouse-IgG, labelled with the same fluorophore (Alexa Fluor 488) was used for specificity-tests of the IgG/anti-IgG interaction.
The immunoassay based on fluorescence was characterized using an optical microscope (Zeiss AXIO inverted fluorescence microscope) working at the wavelengths of 470 nm for excitation of Alexa Fluor 488.
The real time measurement of the resonance broadening after each functionalization step together with the high Q factor (< 105) measured after the IgG/anti-IgG interaction in water, gives a further proof for the method validity.
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