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MicroRNAs (miRNAs) have emerged as a promising class of biomarkers for early detection of various cancers, including ovarian cancer. However, quantifying miRNAs in human blood samples is challenging owing to the issues of sensitivity and specificity. In this study, hsa-miR-200a-3p of the miR-200a sub-family, which is a biomarker of ovarian cancer, was used as the analyte to demonstrate the analytical capability of an integrated biosensing platform using an extremely sensitive surface-enhanced Raman scattering (SERS) nanotag-nanoaggregate-embedded beads (NAEBs), magnetic nanoparticles (MNPs), a pair of highly specific locked nucleic acid (LNA) probes, and a semi-automated paper-based electrowetting-on-dielectric (pEWOD) device to provide labor-less and thorough sample cleanup and recovery. A sandwich approach where NAEBs are modified by one LNA-1 probe and MNPs are modified by another LNA-2 probe was applied. Then, the target analyte miRNA-200a-3p was introduced to form a sandwich nanocomplex through hybridization with the pair of LNA probes. The pEWOD device was used to achieve short cleanup time and good recovery of the nanocomplex, bringing the total analysis time to less than 30 min. The detection limit of this approach can reach 0.26 fM through SERS detection. The versatility of this method without the need for RNA extraction from clinical samples is expected to have good potential in detecting other miRNAs.

Gold nanoparticles (AuNPs) are extensively used in cellular imaging, single-particle tracking, disease diagnosis, studying membrane protein interaction, and drug delivery. Understanding the dynamics of AuNP uptake in live cells is crucial for optimizing their efficacy and safety. Traditional manual methods for quantifying AuNP uptake are time-consuming and subjective, limiting their scalability and accuracy. The available fluorescence-based techniques are limited to photobleaching and photoblinking. Optical microscopy techniques are limited by diffraction limits. Electron microscopy-based imaging techniques are destructive and unsuitable for live cell imaging. Furthermore, the resulting images may contain hundreds of particles with varied intensities, blurring, and substantial occlusion, making it difficult to manually quantify AuNP uptake. To overcome this issue and measure AuNP uptake by live cells, we annotated a dataset of dark-field images of 50 nanometer-radius AuNPs at different incubation durations. Then, to count the number of particles present in a cell, we created a customized multi-column convolutional neural network (MC-CNN). The customized MC-CNN outperformed typical particle counting architectures when compared to spectroscopy-based counting. This will allow researchers to gain a better understanding of AuNP behavior and interactions with cells, paving the way for advancements in nanomedicine, drug delivery, and biomedical research. The code for this paper is available at the following link: https://github.com/Namerlight/LabelFree_AuNP_Quantification.

A portable and non-invasive sensor presents an innovative way to measure inflammation biomarkers in exhaled breath condensate (EBC). This research is focused on developing a miniaturized bipolar electrochemical sensor that can be connected to a smartphone app. This device will be able to detect adding known amounts of nitrite (spikes) to a salt solution and small amounts of nitrite ions in collected real samples in EBC. The sensor was fabricated and tested for its rapid electron transfer capability and ability to detect nitrite ions even at very low concentrations and low real sample levels. In the proposed setup, when the required potential was applied by using a direct power supply, the nitrite ions were oxidized electrocatalytically at amine-functionalized graphene oxide (AGO) decorated with gold nanoparticles on a carbon paper anodic pole. On the other hand, the reduction reaction of Prussian blue occurred at the cathodic pole of the bipolar electrode simultaneously. This strategy led to a change in color from blue to white as a result of the reduction process and the color change is proportional to the concentration of nitrite ions in the analytical solution. The combination of smartphones with the colorimetric method has resulted in a platform for the detection of test strips that is more visual and convenient. The amperometry and voltammetric methods of nitrite detection showed a linear range of up to 1230 μM. The bipolar electrochemical sensor was able to detect the clinically relevant range of nitrite from 0.5 to 85 μM in a buffer with an ultralow detection limit (LOD) of 250 nM (S/N = 3), fast response and excellent selectivity. It was benchmarked by utilizing pre-characterized real EBC samples to differentiate patients with respiratory diseases from healthy volunteers. By tracking the results of nitrite measurements over time, it has become possible to detect trends and changes in an individual's nitrite ion concentration and to potentially identify lung inflammation earlier.

A biosensor is a device that reacts with the analyte to be analyzed, detects its concentration, and generates readable information, which plays an important role in medical diagnosis, detection of physiological indicators, and disease prevention. Nanomaterials have received increasing attention in the fabrication and improvement of biosensors due to their unique physicochemical and optical properties. In this paper, the properties of nanomaterials such as the size effect, optical and electrical properties, and their advantages in the field of biosensing are briefly summarized, and the application of nanomaterials can effectively improve the sensitivity and reduce the detection limit of biosensors. The advantages of commonly used nanomaterials such as gold nanoparticles (AuNPs), carbon nanotubes (CNTs), quantum dots (QDs), graphene, and magnetic nanobeads for biosensor applications are also reviewed. Besides, the two main types of biosensors using nanomaterials involved in their construction and their working principles are described, and the toxicity and biocompatibility of nanomaterials and the future direction of nanomaterial biosensors are discussed.

Bimodal detection facilitates the accurate and reliable detection of cancer biomarkers, which can assist in the early diagnosis of cancer. Herein, S-doped carbon dots (OCDs) with a size of 3 nm and blue emission were synthesized by the hydrothermal treatment of onion extract. The S-doped carbon dots were bioconjugated with an antibody (OCDs@ PSA Ab HRP ) to design a nanoprobe for the detection of prostate specific antigen (PSA), an important serum based prostate cancer biomarker. The detection probe enabled the biomodal assay of PSA via fluorescence immunoassay (FIA) and electrochemical immunoassay (ECIA). In both assays, polyethylenimine stabilized polyaniline nanoparticles (PNPs) were used as the immobilization matrix, which played a major role in widening the linear range of biosensors (0.1 to 100 ng ml -1 for ECIA and 5 to 120 ng ml -1 for FIA). Paper-based and smartphone-integrated fluorescence immuno-array developed using the OCDs@ PSA Ab HRP detection probe provided cost-effective and rapid detection, while the electrochemical immunoassay provided a high sensitivity (7.8 μA ng -1 ml -1 cm -2 ) and low detection limit (38 pg ml -1 ) for PSA detection. The role of OCDs in enhancing the sensor performance was deciphered by carrying out detailed electrochemical studies with HRP enzyme-loaded OCDs. The biosensor was used to detect PSA in human blood serum samples and the results were consistent with conventional techniques. Owing to its analytical properties coupled with simplicity, cost-effectiveness, and portability, the bimodal sensor system has potential for application in clinical analysis.