Electrochemically deposited bimetallic SERS substrate for trace sensing of antibiotics.

Electrochemically deposited bimetallic copper-gold nanoparticles on indium tin oxide (Cu-AuNPs on ITO) glass are demonstrated to be a sensitive and reproducible surface-enhanced Raman scattering (SERS) platform. An optimal signal enhancement with reasonably good degree of homogeneity was obtained by tuning the deposition parameters of the electrochemical setup. For Raman active analytes such as malachite green (MG) and rhodamine 6G (R6G), the developed SERS platform yields a limit of detection (LOD) of 0.75 nM. The usability of the proposed SERS platform has been realized through detection of two important antibiotics namely sulfamethoxazole (SFZ) and tetracycline hydrochloride (TCH) commonly used in egg farms. Furthermore, a machine learning (ML)-based model coupled with a dimensionality reduction technique-principal component analysis (PCA)-has been implemented to classify the targeted analytes in egg samples.

Detection and analysis of rotavirus in clinical stool samples using silver nanoparticle functionalized paper as SERS substrate.

Among the different analytical techniques, surface-enhanced Raman scattering (SERS) approach is a widely used technique for the detection and analysis of various chemicals and biological samples. Present study reports a low-cost, sensitive SERS substrate that has an ability to detect rotavirus in clinical stool samples. The proposed SERS substrate has been fabricated through drop-casting of silver nanoparticles (AgNPs) on a printing-grade paper. Rotavirus particles were extracted from clinical stool samples. The presence of rotavirus antigen in stool samples was confirmed using enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR), and sequencing. The characteristic Raman peaks of rotavirus (RV) particles in solution were found to be significantly enhanced when Raman signals were recorded from the paper-based SERS substrates. Using the proposed SERS substrate, rotavirus samples with concentration as low as 1% could be reliably recorded by the Raman spectrometer. The paper SERS substrate reported herein is an extremely cost-efficient platform and may find applications in other research and clinical laboratories as well.

SERS determination and multivariate classification of antibiotics in chicken meat using gold nanoparticle-decorated electrospun PVA nanofibers.

The fabrication of SERS substrate by gold nanoparticle-decorated polyvinyl alcohol electrospun nanofibers which has been used to detect trace sensing of two widely used poultry antibiotics doxycycline hydrochloride and enrofloxacin is demonstrated. The performance of the backscattered Raman signals from the proposed SERS substrate has been initially evaluated with two standard Raman active compounds namely malachite green and rhodamine-6G. The limit of detection of the proposed substrate is estimated to be 7.32 nM. Following this, the usability of the proposed SERS substrate has been demonstrated through the detection of the aforementioned antibiotics in chicken meat samples. The presence of antibiotics in chicken meat sample has been validated with the standard analytical tool of liquid chromatography-mass spectrometry and the results were compared with the proposed sensing technique. Further, principal component analysis has been performed to classify the antibiotics that are present in the field-collected meat samples.

Gold nanoparticle decorated blu-ray digital versatile disc as a highly reproducible surface-enhanced Raman scattering substrate for detection and analysis of rotavirus RNA in laboratory environment.

Detection and estimation of various biomolecular samples are often required in research and clinical laboratory applications. Present work demonstrates the functioning of a surface-enhanced Raman scattering (SERS) substrate that has been obtained by drop-casting of citrate-reduced gold nanoparticles (AuNPs) of average dimension of 23 nm on a bare blu-ray digital versatile disc (BR-DVD) substrate. The performance of the proposed SERS substrate has been initially evaluated with standard Raman active samples, namely malachite green (MG) and 1,2-bis(4-pyridyl)ethylene (BPE). The designed SERS substrate yields an average enhancement factor of 3.2 × 106 while maintaining reproducibility characteristics as good as 94% over the sensing region of the substrate. The usability of the designed SERS substrate has been demonstrated through the detection and analysis of purified rotavirus double-stranded RNA (dsRNA) samples in the laboratory environment condition. Rotavirus RNA concentrations as low as 10 ng/µL could be detected with the proposed sensing scheme.

Design, fabrication and testing of 3D printed smartphone-based device for collection of intrinsic fluorescence from human cervix.

Fluorescence spectroscopy has the potential to identify discriminatory signatures, crucial for early diagnosis of cervical cancer. We demonstrate here the design, fabrication and testing of a 3D printed smartphone based spectroscopic device. Polarized fluorescence and elastic scattering spectra are captured through the device using a 405 nm laser and a white LED source respectively. The device has been calibrated by comparison of spectra of standard fluorophores (Flavin adenine dinucleotide, fluorescein, rhodamine, and porphyrin) with the corresponding spectra collected from a commercial spectrometer. A few cervical tissue spectra have also been captured for proof of its applicability as a portable, standalone device for the collection of intrinsic fluorescence spectra from human cervix.

Synergy of Adsorption and Plasmonic Photocatalysis in the Au-CeO2 Nanosystem: Experimental Validation and Plasmonic Modeling.

Adsorption-mediated water treatment leaves adsorbents as secondary pollutants in the environment. However, photocatalysis aids in decomposing the contaminant into its nontoxic forms. In this context, we demonstrate an adsorption-photocatalysis pairing in Au-CeO2 nanocomposites for a total methylene blue (MB) removal from water. We synthesized Au-CeO2 through the citrate (cit) reduction method at different Au loading and studied its adsorption capacity with kinetics and thermodynamic models. We observe that the high adsorption capacity of Au-CeO2 is primarily because of the presence of Ce3+ states in CeO2 and citrate ligands on Au NPs. The Ce3+ states interact and transfer their electrons to supported Au NPs, rendering a negative charge over Au. The negatively charged Au surface and the carboxyl (-COO-) group of citrate ligands mediate an electrostatic interaction/adsorption of cationic MB. The total removal of MB is expedited under white light and lasers. A control experiment with Au NPs shows less adsorption-photocatalysis. The size of Au NPs and Au-CeO2 interfacial interaction is responsible for the surface plasmon resonance spectral position at 550-600 nm. Linear sweep voltammetry (LSV) and plasmonic field simulation show surface plasmon-driven photocatalysis in Au-CeO2. LSV shows a 3-fold higher photocurrent density in Au-CeO2 than colloidal Au NPs under white light. The simulated electric field intensity in Au-CeO2 is maximum at SPR excitation and the closest interfacial separation (d = 0 nm). The plasmon-driven photocatalysis in colloidal Au NPs is mainly due to the interaction of hot electrons with the adsorbed MB molecule. Notably, near-field light concentration, hot electrons, and interfacial charge separation are responsible for excellent MB removal in the Au-CeO2 nanosystem. The total MB removal through adsorption-photocatalysis pairing is 99.3% (Au-CeO2), 30.7% (Au NPs), and 13% (CeO2).

An affordable, handheld multimodal microscopic system with onboard cell morphology and counting features on a mobile device.

Microscopes, bright-field (BF) and fluorescence microscopes, in particular, are ubiquitous for clinical diagnostics, cellular and microbiological investigations and in research laboratories. However, the size, cost, fragility and need for skilled personnel to operate these tools restrict their use in resource-limited settings. As an alternative platform, herein, we report a flexible multimodal imaging system that operates in BF and fluorescence modes using a smartphone. Our device utilizes the inbuilt primary camera of phones, and with the aid of easily available optical components, the designed platform is transformed into a high-throughput microscopic device that performs on par with that of a laboratory-grade microscope. The designed platform operates at three different optical magnifications and yields a lateral resolution of 1.21 µm over an acceptable field-of-view (FoV) of diameter ∼4530 µm. The versatility of the device has been demonstrated through imaging of standard microbeads and human blood samples both in BF and fluorescence modes of imaging. Furthermore, the designed imaging platform is equipped with an on-board cell recognition feature which has been obtained through developing a smartphone application for automatic cell counting with high precision.

Immunoinformatics mapping of potential epitopes in SARS-CoV-2 structural proteins.

All approved coronavirus disease 2019 (COVID-19) vaccines in current use are safe, effective, and reduce the risk of severe illness. Although data on the immunological presentation of patients with COVID-19 is limited, increasing experimental evidence supports the significant contribution of B and T cells towards the resolution of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Despite the availability of several COVID-19 vaccines with high efficacy, more effective vaccines are still needed to protect against the new variants of SARS-CoV-2. Employing a comprehensive immunoinformatic prediction algorithm and leveraging the genetic closeness with SARS-CoV, we have predicted potential immune epitopes in the structural proteins of SARS-CoV-2. The S and N proteins of SARS-CoV-2 and SARS-CoVs are main targets of antibody detection and have motivated us to design four multi-epitope vaccines which were based on our predicted B- and T-cell epitopes of SARS-CoV-2 structural proteins. The cardinal epitopes selected for the vaccine constructs are predicted to possess antigenic, non-allergenic, and cytokine-inducing properties. Additionally, some of the predicted epitopes have been experimentally validated in published papers. Furthermore, we used the C-ImmSim server to predict effective immune responses induced by the epitope-based vaccines. Taken together, the immune epitopes predicted in this study provide a platform for future experimental validations which may facilitate the development of effective vaccine candidates and epitope-based serological diagnostic assays.

Turbidimetric analysis of growth kinetics of bacteria in the laboratory environment using smartphone.

For different microbiological and pathological studies, it is often required to monitor the growth of bacteria in a cultured medium in the laboratory environment. UV-VIS spectrophotometer is commonly used to estimate the growth of bacterial cell population by measuring the absorbance at 600 nm over a period of time. Colony-forming unit (CFU) is another approach, which has been routinely performed to estimate the live bacterial cells on semisolid agar plates. Herein, we demonstrate an alternative yet highly reliable sensing platform on a smartphone using which growth kinetics of different bacteria can be reliably monitored. The performance of the proposed smartphone sensor has been compared with the data obtained from OD600 and CFU analysis. A good correlation of bacterial growth rates enumerated based on the proposed smartphone sensor, bench-top spectrophotometer and CFU analysis have been observed under the experimental conditions.

Protein, enzyme and carbohydrate quantification using smartphone through colorimetric digitization technique.

In this paper the utilization of smartphone as a detection platform for colorimetric quantification of biological macromolecules has been demonstrated. Using V-channel of HSV color space, the quantification of BSA protein, catalase enzyme and carbohydrate (using D-glucose) have been successfully investigated. A custom designed android application has been developed for estimating the total concentration of biological macromolecules. The results have been compared with that of a standard spectrophotometer which is generally used for colorimetric quantification in laboratory settings by measuring its absorbance at a specific wavelength. The results obtained with the designed sensor is found to be similar when compared with the spectrophotometer data. The designed sensor is low cost, robust and we envision that it could promote diverse fields of bio-analytical investigations. Schematic illustration of the smartphone sensing mechanism for colorimetric analysis of biomolecular samples.

Low-Cost, Robust, and Field Portable Smartphone Platform Photometric Sensor for Fluoride Level Detection in Drinking Water.

Groundwater is the major source of drinking water for people living in rural areas of India. Pollutants such as fluoride in groundwater may be present in much higher concentration than the permissible limit. Fluoride does not give any visible coloration to water, and hence, no effort is made to remove or reduce the concentration of this chemical present in drinking water. This may lead to a serious health hazard for those people taking groundwater as their primary source of drinking water. Sophisticated laboratory grade tools such as ion selective electrodes (ISE) and portable spectrophotometers are commercially available for in-field detection of fluoride level in drinking water. However, such tools are generally expensive and require expertise to handle. In this paper, we demonstrate the working of a low cost, robust, and field portable smartphone platform fluoride sensor that can detect and analyze fluoride concentration level in drinking water. For development of the proposed sensor, we utilize the ambient light sensor (ALS) of the smartphone as light intensity detector and its LED flash light as an optical source. An android application "FSense" has been developed which can detect and analyze the fluoride concentration level in water samples. The custom developed application can be used for sharing of in-field sensing data from any remote location to the central water quality monitoring station. We envision that the proposed sensing technique could be useful for initiating a fluoride removal program undertaken by governmental and nongovernmental organizations here in India.

Smartphone-based platform optical setup measuring π/256 optical phase difference in an interference process.

Utilizing the camera of a smartphone and simple laboratory optical components, we demonstrate an optical technique that measures an optical phase difference (OPD) of π/256 in an interference process. We develop a compact optical setup for viewing circular interference fringe patterns through the camera of the smartphone. By introducing OPD between the interfering beams, variation in fringe pattern is recorded using the smartphone camera. We envision that the proposed optical setup could emerge as an ultrasensitive optical tool for measurement of inclination of a given surface.

Label-free biodetection using a smartphone.

Utilizing its integrated camera as a spectrometer, we demonstrate the use of a smartphone as the detection instrument for a label-free photonic crystal biosensor. A custom-designed cradle holds the smartphone in fixed alignment with optical components, allowing for accurate and repeatable measurements of shifts in the resonant wavelength of the sensor. Externally provided broadband light incident upon an entrance pinhole is subsequently collimated and linearly polarized before passing through the biosensor, which resonantly reflects only a narrow band of wavelengths. A diffraction grating spreads the remaining wavelengths over the camera's pixels to display a high resolution transmission spectrum. The photonic crystal biosensor is fabricated on a plastic substrate and attached to a standard glass microscope slide that can easily be removed and replaced within the optical path. A custom software app was developed to convert the camera images into the photonic crystal transmission spectrum in the visible wavelength range, including curve-fitting analysis that computes the photonic crystal resonant wavelength with 0.009 nm accuracy. We demonstrate the functionality of the system through detection of an immobilized protein monolayer, and selective detection of concentration-dependent antibody binding to a functionalized photonic crystal. We envision the capability for an inexpensive, handheld biosensor instrument with web connectivity to enable point-of-care sensing in environments that have not been practical previously.

Fiber-optic liquid level sensor based on coupling optical path length variation.

The concept for a new and simple fiber-optic liquid level sensor is presented and experimental results are shown to demonstrate the principle. The sensing principle is based on light intensity modulation when rising and falling mode of liquid level causes coupling optical path distance variation between two optical fibers. Near continuous mode of liquid level variation could be monitored with resolution as low as 1 mm can be measured in the length scale of 25 cm.