ABSTRACT
A label-free sensor, based on the combination of silicon photonic bandgap (PBG) structures with immobilized molecular beacon (MB) probes, is experimentally developed. Complementary target oligonucleotides are specifically recognized through hybridization with the MB probes on the surface of the sensing structure. This combination of PBG sensing structures and MB probes demonstrates an extremely high sensitivity without the need for complex PCR-based amplification or labelling methods.
ABSTRACT
An experimental study of the influence of the conformational change suffered by molecular beacon (MB) probes-upon the biorecognition of nucleic acid target oligonucleotides over evanescent wave photonic sensors-is reported. To this end, high sensitivity photonic sensors based on silicon photonic bandgap (PBG) structures were used, where the MB probes were immobilized via their 5' termination. Those MBs incorporate a biotin moiety close to their 3' termination in order to selectively bind a streptavidin molecule to them. The different photonic sensing responses obtained toward the target oligonucleotide detection, when the streptavidin molecule was bound to the MB probes or not, demonstrate the conformational change suffered by the MB upon hybridization, which promotes the displacement of the streptavidin molecule away from the surface of the photonic sensing structure.
Subject(s)
Nucleic Acid Conformation , Oligonucleotide Probes/chemistry , Optical Devices , Photons , Electrochemistry , Models, Molecular , Nucleic Acid HybridizationABSTRACT
Porous silicon seems to be an appropriate material platform for the development of high-sensitivity and low-cost optical sensors, as their porous nature increases the interaction with the target substances, and their fabrication process is very simple and inexpensive. In this paper, we present the experimental development of a porous silicon microcavity sensor and its use for real-time in-flow sensing application. A high-sensitivity configuration was designed and then fabricated, by electrochemically etching a silicon wafer. Refractive index sensing experiments were realized by flowing several dilutions with decreasing refractive indices, and measuring the spectral shift in real-time. The porous silicon microcavity sensor showed a very linear response over a wide refractive index range, with a sensitivity around 1000 nm/refractive index unit (RIU), which allowed us to directly detect refractive index variations in the 10-7 RIU range.