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A plasmonic gold nanofilm-based microfluidic chip for rapid and inexpensive droplet-based photonic PCR.

ABSTRACT: Polymerase chain reaction (PCR) is a powerful tool for nucleic acid amplification and quantification. However, long thermocycling time is a major limitation of the commercial PCR devices in the point-of-care (POC). Herein, we have developed a rapid droplet-based photonic PCR (dpPCR) system, including a gold (Au) nanofilm-based microfluidic chip and a plasmonic photothermal cycler. The chip is fabricated by adding mineral oil to uncured polydimethylsiloxane (PDMS) to suppress droplet evaporation in PDMS microfluidic chips during PCR thermocycling. A PDMS to gold bonding technique using a double-sided adhesive tape is applied to enhance the bonding strength between the oil-added PDMS and the gold nanofilm. Moreover, the gold nanofilm excited by two light-emitting diodes (LEDs) from the top and bottom sides of the chip provides fast heating of the PCR sample to 230 °C within 100 s. Such a design enables 30 thermal cycles from 60 to 95 °C within 13 min with the average heating and cooling rates of 7.37 ± 0.27 °C/s and 1.91 ± 0.03 °C/s, respectively. The experimental results demonstrate successful PCR amplification of the alcohol oxidase (AOX) gene using the rapid plasmonic photothermal cycler and exhibit the great performance of the microfluidic chip for droplet-based PCR.

SUBMITTER: Jalili A

PROVIDER: S-EPMC8639772 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Hemoglobin is an iron carrying protein in erythrocytes and also an essential element to transfer oxygen from the lungs to the tissues. Abnormalities in hemoglobin concentration are closely correlated with health status and many diseases, including thalassemia, anemia, leukemia, heart disease, and excessive loss of blood. Particularly in resource-constrained settings existing blood analyzers are not readily applicable due to the need for high-level instrumentation and skilled personnel, thereby inexpensive, easy-to-use, and reliable detection methods are needed. Herein, a molecular fingerprints of hemoglobin on a nanofilm chip was obtained for real-time, sensitive, and selective hemoglobin detection using a surface plasmon resonance system. Briefly, through the photopolymerization technique, a template (hemoglobin) was imprinted on a monomeric (acrylamide) nanofilm on-chip using a cross-linker (methylenebisacrylamide) and an initiator-activator pair (ammonium persulfate-tetramethylethylenediamine). The molecularly imprinted nanofilm on-chip was characterized by atomic force microscopy and ellipsometry, followed by benchmarking detection performance of hemoglobin concentrations from 0.0005 mg mL-1 to 1.0 mg mL-1. Theoretical calculations and real-time detection implied that the molecularly imprinted nanofilm on-chip was able to detect as little as 0.00035 mg mL-1 of hemoglobin. In addition, the experimental results of hemoglobin detection on the chip well-fitted with the Langmuir adsorption isotherm model with high correlation coefficient (0.99) and association and dissociation coefficients (39.1 mL mg-1 and 0.03 mg mL-1) suggesting a monolayer binding characteristic. Assessments on selectivity, reusability and storage stability indicated that the presented chip is an alternative approach to current hemoglobin-targeted assays in low-resource regions, as well as antibody-based detection procedures in the field. In the future, this molecularly imprinted nanofilm on-chip can easily be integrated with portable plasmonic detectors, improving its access to these regions, as well as it can be tailored to detect other proteins and biomarkers.

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A plasmonic gold nanofilm-based microfluidic chip for rapid and inexpensive droplet-based photonic PCR.

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