An aptamer-based paper microfluidic device for the colorimetric determination of cocaine.

A method utilizing paper microfluidics coupled with gold nanoparticles and two anticocaine aptamers has been developed to detect seized cocaine samples. The ready-to-use format involves the use of a paper strip that produces a color change resulting from the salt-induced aggregation of gold nanoparticles producing a visible color change indicating the presence of the drug. This format is specific for the detection of cocaine. The visual LOD for the method was 2.5 µg and the camera based LOD was 2.36 µg. The operation of the device is easy and rapid, and does not require extensive training or instrumentation. All of the materials utilized in the device are safe and environmental friendly. This device should prove a useful tool for the screening of forensic samples.

A Fully Integrated Paper-Microfluidic Electrochemical Device for Simultaneous Analysis of Physiologic Blood Ions.

A fully integrated paper microfluidic electrochemical device equipped with three different cation permeable films is developed to determine blood ions (Cl-, Na⁺, K⁺, and Ca2+) at a time. These blood ions that are normally dissolved in the real human blood stream are essential for cell metabolisms and homeostasis in the human body. Abnormal concentration of blood ions causes many serious disorders. The optimized microfluidic device working without any external power source can directly and effectively separate human blood components, and subsequently detect a specific blood ion with minimized interference. The measured sensitivity to Cl-, K⁺, Na⁺, and Ca2+ are -47.71, 45.97, 51.06, and 19.46 in mV decade-1, respectively. Potentiometric responses of the microfluidic devices to blood serum samples are in the normal ranges of each cation, and comparable with responses from the commercial blood ion analyzer Abbott i-Stat.

Paper based microfluidic platform for single-step detection of mesenchymal stromal cells secreted VEGF.

Low cost and user-friendly paper microfluidic devices, combined with DNA-based biosensors with binding capacities for specific molecules, have been proposed for the developing of novel platforms that ease and speed-up the process of cell secretion monitoring. In this work, we present the first cellulose microfluidic paper-based analytical device for the single-step detection of cell secreted Vascular Endothelial Growth Factor through a self-reporting Structure Switching Signaling Aptamer. A three-part Structure Switching Signaling Aptamer was designed with an aptameric sequence specific for VEGF, which provides a quantifiable fluorescent signal through the displacement of a quencher upon VEGF recognition. The VEGF biosensor was integrated in cellulose paper, enabling the homogenous distribution of the sensor in the paper substrate and the detection of as low as 0.34 ng of VEGF in 30 min through fluorescence intensity analysis. As a proof-of-concept, the biosensor was incorporated in a microfluidic paper-based analytical device format containing a VEGF detection zone and a control zone, which was applied for the detection of cell secreted VEGF in the supernatant of mesenchymal stem cells culture plates, demonstrating its potential use in cell biology research.

DNA-copper hybrid nanoflowers as efficient laccase mimics for colorimetric detection of phenolic compounds in paper microfluidic devices.

Laccases are important multicopper oxidases that are involved in many biotechnological processes; however, they suffer from poor stability as well as high cost for production/purification. Herein, we found that DNA-copper hybrid nanoflowers, prepared via simple self-assembly of DNA and copper ions, exhibit an intrinsic laccase-mimicking activity, which is significantly higher than that of control materials formed in the absence of DNA. Upon testing all four nucleobases, we found that hybrid nanoflowers composed of guanine-rich ssDNA and copper phosphate (GNFs) showed the highest catalytic activity, presumably due to the affirmative coordination between guanine and copper ions. At the same mass concentration, GNFs had similar Km but 3.5-fold higher Vmax compared with those of free laccase, and furthermore, they exhibited significantly-enhanced stability in ranges of pH, temperature, ionic strength, and incubation period of time. Based on these advantageous features, GNFs were applied to paper microfluidic devices for colorimetric detection of diverse phenolic compounds such as dopamine, catechol, and hydroquinone. In the presence of phenolic compounds, GNFs catalyzed their oxidation to react with 4-aminoantipyrine for producing a colored adduct, which was conveniently quantified from an image acquired using a conventional smartphone with ImageJ software. Besides, GNFs successfully catalyzed the decolorization of neutral red dye much faster than free laccase. This work will facilitate the development of nanoflower-type nanozymes for a wide range of applications in biosensors and bioremediation.

Real time monitoring of glucose in whole blood by smartphone.

A combined thread-paper microfluidic device (µTPAD) is presented for the determination of glucose in blood. The device is designed to include all the analytical operations needed: red blood cell separation, conditioning, enzymatic recognition, and colorimetric transduction. The signal is captured with a smartphone or tablet working in video mode and processed by custom Android-based software in real-time. The automatic detection of the region of interest on the thread allows for the use of either initial rate or equilibrium signal as analytical parameters. The time needed for analysis is 12 s using initial rate, and 100 s using the equilibrium measurement with a LOD of 48 µM and 12 µM, respectively, and a precision around 7%. The µTPAD allows a rapid determination of glucose in real samples using only 3 µL of whole blood.

Graphene nanocomposites modified electrochemical aptamer sensor for rapid and highly sensitive detection of prostate specific antigen.

Prostate specific antigen (PSA) is a widely used marker for the diagnosis of prostate cancer, and the increasing attention has been attracted on the development of rapid assay using biosensing technology. However, it remains challenging for the sensitive and selective detection of PSA in clinical samples. Here, we report a label-free microfluidic paper-based analytical device for highly sensitive electrochemical detection of PSA. The paper device was fabricated with wax printing to generate hydrophobic and hydrophilic layers for the construction of microfluidic channel, followed by screen-printing of three electrodes including working, counter and reference electrode. Gold nanoparticles (AuNPs)/reduced graphene oxide (rGO)/thionine (THI) nano composites were synthesized and characterized, which were coated onto working electrodes for the immobilization of DNA aptamer probe. THI servers as the electrochemical mediator to transduce the biological recognition between DNA aptamer and PSA, and the excellent conductivity of AuNPs and rGO also play a significant role of electron transfer, leading to a sensitive detection for PSA, able to detect PSA as low as 10 pg mL-1, with a linear range from 0.05 to 200 ng mL-1. We demonstrated that our electrochemical sensor for the detection of clinical serum samples, indicating that our sensor would provide a new platform for low cost, sensitive and point-of-care diagnosis of prostate cancer.