SERS-active metal-dielectric nanostructures integrated in microfluidic devices for label-free quantitative detection of miRNA

Author:

Novara Chiara1234,Chiadò Alessandro1234,Paccotti Niccolò1234,Catuogno Silvia564,Esposito Carla Lucia564,Condorelli Gerolama56478,De Franciscis Vittorio564,Geobaldo Francesco1234,Rivolo Paola1234,Giorgis Fabrizio1234

Affiliation:

1. Department of Applied Science and Technology

2. Politecnico di Torino

3. Turin

4. Italy

5. Institute of Endocrinology and Experimental Oncology of Italian National Research Council

6. Naples

7. Department of Molecular Medicine and Medical Biotechnology

8. “Federico II” University of Naples

Abstract

In this work, SERS-based microfluidic PDMS chips integrating silver-coated porous silicon membranes were used for the detection and quantitation of microRNAs (miRNAs), which consist of short regulatory non-coding RNA sequences typically over- or under-expressed in connection with several diseases such as oncogenesis. In detail, metal–dielectric nanostructures which provide noticeable Raman enhancements were functionalized according to a biological protocol, adapted and optimized from an enzyme-linked immunosorbent assay (ELISA), for the detection of miR-222. Two sets of experiments based on different approaches were designed and performed, yielding a critical comparison. In the first one, the labelled target miRNA is revealed through hybridization to a complementary thiolated DNA probe, immobilized on the silver nanoparticles. In the second one, the probe is halved into shorter strands (half1 and half2) that interact with the complementary miRNA in two steps of hybridization. Such an approach, taking advantage of the Raman labelling of half2, provides a label-free analysis of the target. After suitable optimisation of the procedures, two calibration curves allowing quantitative measurements were obtained and compared on the basis of the SERS maps acquired on the samples loaded with several miRNA concentrations. The selectivity of the two-step assay was confirmed by the detection of target miR-222 mixed with different synthetic oligos, simulating the hybridization interference coming from similar sequences in real biological samples. Finally, that protocol was applied to the analysis of miR-222 in cellular extracts using an optofluidic multichamber biosensor, confirming the potentialities of SERS-based microfluidics for early-cancer diagnosis.

Publisher

Royal Society of Chemistry (RSC)

Subject

Physical and Theoretical Chemistry

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