A Fluorescent Biosensor for Sensitive Detection of Salmonella Typhimurium Using Low-Gradient Magnetic Field and Deep Learning via Faster Region-Based Convolutional Neural Network

In this study, a fluorescent biosensor was developed for the sensitive detection of Salmonella typhimurium using a low-gradient magnetic field and deep learning via faster region-based convolutional neural networks (R-CNN) to recognize the fluorescent spots on the bacterial cells. First, magnetic nanobeads (MNBs) coated with capture antibodies were used to separate target bacteria from the sample background, resulting in the formation of magnetic bacteria. Then, fluorescein isothiocyanate fluorescent microspheres (FITC-FMs) modified with detection antibodies were used to label the magnetic bacteria, resulting in the formation of fluorescent bacteria. After the fluorescent bacteria were attracted against the bottom of an ELISA well using a low-gradient magnetic field, resulting in the conversion from a three-dimensional (spatial) distribution of the fluorescent bacteria to a two-dimensional (planar) distribution, the images of the fluorescent bacteria were finally collected using a high-resolution fluorescence microscope and processed using the faster R-CNN algorithm to calculate the number of the fluorescent spots for the determination of target bacteria. Under the optimal conditions, this biosensor was able to quantitatively detect Salmonella typhimurium from 6.9 × 101 to 1.1 × 103 CFU/mL within 2.5 h with the lower detection limit of 55 CFU/mL. The fluorescent biosensor has the potential to simultaneously detect multiple types of foodborne bacteria using MNBs coated with their capture antibodies and different fluorescent microspheres modified with their detection antibodies.

Do plexiglass barriers reduce the risk for transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)?

Barriers are commonly installed in workplace situations where physical distancing cannot be maintained, but their effectiveness in decreasing viral transmission is unknown. In simulations, physical barriers with no openings were effective in reducing contamination with an aerosolized benign virus or fluorescent microspheres, but barriers with openings were not.

Portable, Rapid, and Sensitive Time-Resolved Fluorescence Immunochromatography for On-Site Detection of Dexamethasone in Milk and Pork

Dexamethasone (DEX) is widely used because of its anti-inflammatory, anti-endotoxin, anti-shock, and stress-enhancing response activities. It can increase the risk of diabetes and hypertension if it is abused or used improperly. However, there is a lack of sensitive and rapid screening methods for DEX in food. In this study, a time-resolved fluorescent microspheres immunochromatographic assay (TRFM-ICA) integrated with a portable fluorescence reader was developed for the quantitative detection of DEX in milk and pork. The cut-off values of the TRFM-ICA were 0.25 ng/mL and 0.7 µg/kg, respectively. The limits of quantitation (LOQs) were 0.003 ng/mL and 0.062 µg/kg, respectively. The recovery rates were 80.0–106.7%, and 78.6–83.6%, respectively, with the coefficients of variation ranging 6.3–12.5%, and 7.5–10.3%, respectively. A parallel experiment for 20 milk and 10 pork samples with LC-MS/MS was carried out to confirm the performance of the on-site application of the developed TRFM-ICA. The results of the two methods are basically the same; the correlation (R2) was >0.98. The establishment of TRFM-ICA will provide a new sensitive and efficient technical support for the rapid screening of DEX in food.

Microplastics and phagotrophic soil protists: evidence of ingestion

Microplastics (MPs) can now be found in all the Earth’s biomes, thereby representing a global change phenomenon with largely unknown consequences for biodiversity and ecosystem functioning. Soil protists are eukaryotic, primarily single celled organisms that play important roles in the soil food web. Microplastics have been shown to affect protist populations in freshwater and marine environments, yet the interactions between soil protists and MPs remains largely unknown. Here we examined whether phagotrophic soil protists can ingest MPs and experience declines in abundance. We exposed protists to soil treatments with different concentrations of MPs using commercial polymer fluorescent microspheres and used fluorescence microscopy to find evidence of MP ingestion. In addition, we quantified the total number of active phagotrophic protists over time. We show that most soil protists (>75% individuals) can readily ingest and keep MP within their food vacuoles, even at relatively small MP concentrations (0.1% w/w). There was a trend for higher prevalence of ingestion and for declines in protist abundance at the highest concentration of MPs (1% w/w). However, more data are necessary to further ascertain cause-effect relationships. This is the first report indicating that soil protists can play an important role in the transport and uptake of MPs in the soil food web.

Synthesis and experimental study of liquid dispersions of magnetic fluorescent polystyrene microspheres

Multiplex microsphere-based immunofluorescence assay is a reliable, accurate, and highly sensitive method for the detection of various biomolecules. However, for the moment, the wide application of the method in clinical practice is prevented by the high cost of reagents for analysis - magnetic spectrally encoded microspheres. Therefore, an urgent task is the development of new methods for the synthesis of microspheres with the required properties. The aim of this study was the creation of new magnetic fluorescent microspheres suitable for use in multiplex immunoassay.Samples of magnetic fluorescent polystyrene microspheres were synthesized by dispersion polymerization and two-stage swelling methods. Experimental studies of geometric parameters, fluorescence, magnetic properties of the synthesized microspheres have been carried out.The results of the studies have shown that microspheres synthesized by dispersion polymerization are promising for the use in immunofluorescence analysis. The obtained results can be used for the development of new diagnostic multiplex test systems based on spectrally encoded microspheres.