M.F. Laguna

Label-free biosensing by means of periodic lattices of high aspect ratio SU-8 nano-pillars

We developed biophotonic sensing arrays of 60x60 microm(2) made of periodic lattices of high aspect ratio SU-8 nano-pillars in order to demonstrate their capability for label-free molecule detection, as well as the sensitivity enhancement in comparison with a single layer of SU-8. The biophotonic sensing arrays, that we call BICELLs (Biophotonic sensing cells), are interrogated vertically by using micron spot size Fourier transform visible and IR spectrometry (FT-VIS-IR). We monitored the surface immobilization of bovine serum albumin (BSA) antigen and anti-BSA antibody (aBSA) recognition. The bioassay exhibits a limit of detection (LOD) in the order of 2 ng/ml limited by the wavenumber uncertainty during the interrogation process. We also estimated and compared the theoretical biolayer thickness with previous results.

Bio-Photonic Sensing Cells over transparent substrates for anti-gestrinone antibodies biosensing.

In a previous work we introduced the term Bio-Photonic Sensing Cells (BICELLs), referred to periodic networks of nano-pillar suitable for biosensing when are vertically interrogated. In this article, we demonstrate the biosensing capabilities of a type of micrometric size BICELLs made of SU-8 nano-pillars fabricated over transparent substrates. We verify the biochips functionality comparing the theoretical simulations with the experimental results when are optically interrogated in transmission. We also demonstrate a sensitivity enhancement by reducing the pitch among nano-pillars from 800 to 700 nm. Thus, the Limit of Detection achievable in these types of BICELLs is in the order of 64 pg/mL for 700 nm in pitch among nano-pillars in comparison with 292 pg/mL for 800 nm in pitch when are interrogated by Fourier Transform Visible and Infrared Spectrometry. The experiments exhibited a good reproducibility with a relative standard deviation of 0.29% measured within 8 days for a specific concentration. Finally, BICELLs functionality was tested in real conditions with unpurified rabbit serum for detecting anti-gestrinone antibodies, demonstrating the high performance of this type of BICELLs to detect specific antibodies having immobilized the suitable bioreceptors onto the sensing surface.

Silicon nanopillar arrays with SiO2 overlayer for biosensing application

We present the fabrication of silicon dioxide (SiO2) coated silicon nanopillar array structures and demonstrate their application as sensitive optical biosensors. Colloidal lithography, plasma dry etching and deposition processes are used to fabricate SiO2 coated Si nanopillar arrays with two different diameters and periods. Proof of concept bio recognition experiments are carried out with the bovine serum albumin (BSA)/antiBSA model system using Fourier transform visible and IR spectrometry (FT-VIS-IR) in reflection mode. A limit of detection (LoD) value of 5.2 ng/ml is estimated taking in to account the wavenumber uncertainty in the measurements.

Bulk sensing performance comparison between silicon dioxide and resonant high aspect ratio nanopillars arrays fabricated by means of interference lithography

In this work we present the refractive index sensing performance comparison between resonant (R-NPs) and silicon dioxide (SiO2-NPs) high-aspect ratio nano-pillars arrays. Both arrays have been fabricated by laser interference lithography and reactive ion etching. The R-NPs are made by a multilayer of silicon oxide and silicon nitride distributed to act as a vertical resonant cavity with two Bragg reflectors. Several chips containing eight periodic arrays of R-NPs and SiO2-NPs were implemented following the presented fabrication process, having a height in the order of 2.5 μm, a diameter in the order of 200 nm, different pitches and aspect ratio up to 9.8. Finally, the optical responses of these arrays were measured by infiltration of fluids with different refractive indexes. The main conclusion is that sensitivity obtained for the R-NPs is more than two times higher in comparison with the SiO2-NPs sensitivity (3724 cm−1/RIU and 1652 cm−1/RIU, respectively).

Arrays of resonant nanopillars for biochemical sensing

In this Letter, we demonstrate for the first time the experimental capability for the biochemical sensing of resonant nanopillars (RNPs) arrays. These arrays are fabricated over a glass substrate and are optically integrated from the backside of this substrate. The reflectivity profiles of the RNPs arrays are measured by infiltrating different ethanol fractions in water in order to evaluate the optical response for the different refractive indexes, which range from 1.330 to 1.342. A linear fit of the resonant modes shift is observed as a function of the bulk refractive index of the liquid infiltrated. For the type of transducer analyzed, a relative sensitivity of 10017  cm(-1)/Refractive Index Unit (RIU) is achieved, allowing us to reach a competitive Limit of Detection (LoD) in the order of 1×10(-5)  RIU.

Optimization of Dengue Immunoassay by Label-Free Interferometric Optical Detection Method

In this communication we report a direct immunoassay for detecting dengue virus by means of a label-free interferometric optical detection method. We also demonstrate how we can optimize this sensing response by adding a blocking step able to significantly enhance the optical sensing response. The blocking reagent used for this optimization is a dry milk diluted in phosphate buffered saline. The recognition curve of dengue virus over the proposed surface sensor demonstrates the capacity of this method to be applied in Point of Care technology.

Description of an Advantageous Optical Label-Free Biosensing Interferometric Read-Out Method to Measure Biological Species

In this article we report a new, simple, and reliable optical read-out detection method able to assess Rotavirus present in human sera as well as in the viral pollution sources. It is based on the interference of two interferometers used as biophotonic transducers. The method significantly improves the optical label-free biosensing response measuring both, the concentration of the AgR and its corresponding size. Two different immunoassays were carried out: Bovine Serum Albumin (BSA), and the recognition by its antibody (anti-BSA); and Rotavirus (AgR) and the recognition by its antibody (anti-AgR). In the cases studied, and using as model interferometer a simple Fabry-Perot transducer, we demonstrate a biosensing enhancement of two orders of magnitude in the Limit of Detection (LoD). In fact, this read-out optical method may have significant implications to enhance other optical label-free photonic transducers reported in the scientific literature.

Biomolecular Interaction Analysis of Gestrinone-anti-Gestrinone Using Arrays of High Aspect Ratio SU-8 Nanopillars

In this paper, label-free biosensing for antibody screening by periodic lattices of high-aspect ratio SU-8 nano-pillars (BICELLs) is presented. As a demonstration, the determination of anti-gestrinone antibodies from whole rabbit serum is carried out, and for the first time, the dissociation constant (KD = 6 nM) of antigen-antibody recognition process is calculated using this sensing system. After gestrinone antigen immobilization on the BICELLs, the immunorecognition was performed. The cells were interrogated vertically by using micron spot size Fourier transform visible and IR spectrometry (FT-VIS-IR), and the dip wavenumber shift was monitored. The biosensing assay exhibited good reproducibility and sensitivity (LOD = 0.75 ng/mL).

Resonant nanopillars arrays for label-free biosensing

In our previous work we demonstrated for the first time, to the best of our knowledge, the experimental capability of resonant nanopillars (R-NP) arrays as biochemical transducers. In this Letter, we provide evidence of the capability and suitability of R-NP arrays on a chip to function as label-free optical multiplexed biosensors. R-NP are based on Si3N4/SiO2 Bragg reflectors with a cavity of SiO2. In order to demonstrate the biosensing performance, R-NP were biofunctionalized by the immobilization of IgG antibodies acting as a bioreceptor. This immobilization was carried out through the silanization of the pillars sensing surface with APTMS (3-aminopropyltrimethoxysilane). R-NP were integrated in eight different sensing arrays on a quartz surface chip. An optical fiber bundle monitored each sensing array vertically and independently after each biofunctionalization step, and subsequently after every recognition event of increasing concentrations of anti-IgGs. The results report a novel multiplexed optical biosensor made of eight sensing arrays on a chip with promising performance and yield.

Development of a versatile biotinylated material based on SU-8

The negative epoxy-based SU-8 photoresist has a wide variety of applications within the semiconductor industry, photonics and lab-on-a-chip devices, and it is emerging as an alternative to silicon-based devices for sensing purposes. In the present work, biotinylation of the SU-8 polymer surface promoted by light is reported. As a result, a novel, effective, and low-cost material, focusing on the immobilization of bioreceptors and consequent biosensing, is developed. This material allows the spatial discrimination depending on the irradiation of desired areas. The most salient feature is that the photobiotin may be directly incorporated into the SU-8 curing process, consequently reducing time and cost. The potential use of this substrate is demonstrated by the immunoanalytical detection of the synthetic steroid gestrinone, showing excellent performances. Moreover, the naked eye biodetection due to the transparent SU-8 substrate, and simple instrumental quantification are additional advantages.