Rapid and Sensitive Quantification of Bacterial Viability Using Ratiometric Fluorescence Sensing.

Bacterial viability assessment plays an important role in food-borne pathogen detection and antimicrobial drug development. Here, we first used GelRed as a DNA-binding stain for a bacterial viability assessment. It was found that live bacteria were able to exclude GelRed, which however could easily penetrate dead ones and be absorbed nonspecifically on the bacterial periplasm.

Gelatin nanocarriers assembled by a self-immolative cross-linker for targeted cancer therapy.

With a number of outstanding properties, gelatin is an ideal candidate for assembling nanoplatforms in biomedical applications. Generally, gelatin nanocarriers are cross-linked by aldehydes to improve their stability in water solution. However, aldehydes could cause multiple toxicities and their cross-linking products are uncontrollable.

Secretory production and characterization of a highly effective chitosanase from Streptomyces coelicolor A3(2) M145 in Pichia pastoris.

In this study, a glycoside hydrolase family 46 chitosanase from Streptomyces coelicolor A3(2) M145 was firstly cloned and expressed in Pichia pastoris GS115 (P. pastoris GS115). The recombinant enzyme (CsnA) showed maximal activity at pH 6.

Transformation of proteins into reproductive DNA templates for sensitive quantification of PSA.

A switch DNA template was designed to transform proteins into linear DNA strands of 97 nt. The linear DNA was subjected to quantitative polymerase chain reaction (qPCR) to determine the initial copy number, which correlated positively with the protein concentration. A restriction endonuclease was used to remove background amplification.

Sensitive detection of aflatoxin B1 in foods by aptasensing-based qPCR.

In this study, a reproductive switch DNA template was designed using aptasensing principles for the accurate quantification of aflatoxins. The template transformed the aflatoxin molecule into linear DNA of 102 nt. The linear DNA was subjected to a quantitative polymerase chain reaction (qPCR) to determine its initial copy number, which was positively correlated with the aflatoxin concentration.

No-cost ballpoint pen dispenser for lateral flow assays.

We have developed a no-cost, lightweight, human-powered dispenser using an empty ballpoint pen. Used in lateral flow assays, this dispenser restricts antibody deposition to narrow zones, allowing freehand drawing of test and control lines. The lines can be drawn in widths ranging from 0.

Reversible cross-linking of gelatin by a disulphide-containing bis-succinimide for tunable degradation and release.

Generally, gelatin was irreversibly cross-linked by chemical reagents to improve its water-resistance. However, few chemical reagents meet both the requirements of high cross-inking efficiency and tunable degradation. Here a reversible cross-linker, disulphide-containing bis-succinimide, was synthesized and used to control the cross-linking and degradation of edible gelatin film.

Intrafibrillar mineralization of type I collagen by micelle-loaded amorphous calcium phosphate nanoparticles.

Mineralization of type I collagen fibrils is highly desired for artificial bone preparation and teeth repairing. Generally, amorphous calcium phosphate (ACP) combined with non-collagenous protein analogue (NCPA) were used for biomimetic remineralization of collagen fibrils. However, the ACP was likely to aggregate to form larger particles that could not infiltrate into the gaps of the collagen for intrafibrillar mineralization, and the poor storage stability of ACP has challenged its practical applications.

Bridge-DNA synthesis triggered by an allosteric aptamer for the colorimetric detection of pathogenic bacteria.

Rapid and sensitive quantification of pathogenic bacteria is highly desired for environmental health supervision and food safety control. Yet, the amplification and detection of bacteria with a concentration lower than 10 cfu mL remains a great challenge. Here, we combined an allosteric aptamer (AAP) with a gold nanoparticle (AuNP) for assembling a bridge-DNA synthesis system (named as AuNP-BDS) to amplify the bacterial signals.

Preparation, characteristics and cytotoxicity of green synthesized selenium nanoparticles using Paenibacillus motobuensis LY5201 isolated from the local specialty food of longevity area.

Selenium is an essential micronutrient element. For the extremely biotoxic of selenite, Selenium nanoparticles (SeNPs) is gaining increasing interest. In this work, a selenium-enriched strain with highly selenite-resistant (up to 173 mmol/L) was isolated from the local specialty food of longevity area and identified as Paenibacillus motobuensis (P.

On-bead DNA synthesis triggered by allosteric probe for detection of carcinoembryonic antigen.

Sensitive quantification of protein biomarkers is highly desired for clinical diagnosis and treatment. Yet, unlike DNA/RNA which can be greatly amplified by PCR/RT-PCR, the amplification and detection of trace amount of proteins remain a great challenge. Here, we combined allosteric probe (AP) with magnetic bead (MB) for assembling an on-bead DNA synthesis system (named as APMB) to amplify protein signals.

Using a safe and effective fixative to improve the immunofluorescence staining of bacteria.

The emerging and development of green chemistry has once again drawn the researchers' attention to eliminating the use and generation of hazardous materials. Here we report the use of a safe and effective fixative, chlorine dioxide (ClO), instead of traditional hazardous fixatives for the cross-linking of cellular proteins to improve immunofluorescence staining of bacteria. The concentration of ClOneeded for 100% fixation is 50g ml, which is much lower than that of traditional fixatives (1000-10000g ml).

Assembly of "carrier free" enzymatic nano-reporters for improved ELISA.

Enzyme-linked immunosorbent assay (ELISA) is an economic and easy operation technique that has been widely used for the detection of protein in industry. However, the low loading capacity of the enzyme reporter has contributed to the low sensitivity of traditional ELISA, and the cross-linking procedures of the enzyme-labeled antibody in ELISA methods can lead to the inactivation of the enzyme, which will further decrease the sensitivity. To address this issue, herein we fabricated "carrier-free" nanoparticles to obtain a horseradish peroxidase (HRP) labelled reporter with a high HRP loading capacity.

Rapid quantification of pathogenic Salmonella Typhimurium and total bacteria in eggs by nano-flow cytometry.

Rapid quantification of pathogenic Salmonella Typhimurium (S. Typhimurium) and total bacteria in eggs is highly desired for food safety control. However, the complexity of egg matrix presents a significant challenge for sensitive detection of bacteria.

Label-Free Detection of Bacteria in Fruit Juice by Nano-Flow Cytometry.

Rapid quantification of microbial contamination in fruit juice is highly desired for food safety control. Yet, the complex sample matrix and the diversity of bacterial contaminants present a great challenge. Employing a laboratory-built nano-flow cytometer (nFCM), here we report the development of a label-free approach for the detection of bacteria population in fruit juice.

Deciphering the Antitoxin-Regulated Bacterial Stress Response via Single-Cell Analysis.

Bacterial toxin-antitoxin (TA) systems, which are diverse and widespread among prokaryotes, are responsible for tolerance to drugs and environmental stresses. However, the low abundance of toxin and antitoxin proteins renders their quantitative measurement in single bacteria challenging. Employing a laboratory-built nano-flow cytometer (nFCM) to monitor a tetracysteine (TC)-tagged TA system labeled with the biarsenical dye FlAsH, we here report the development of a sensitive method that enables the detection of basal-level expression of antitoxin.

Flow Cytometric Analysis of Nanoscale Biological Particles and Organelles.

Analysis of nanoscale biological particles and organelles (BPOs) at the single-particle level is fundamental to the in-depth study of biosciences. Flow cytometry is a versatile technique that has been well-established for the analysis of eukaryotic cells, yet conventional flow cytometry can hardly meet the sensitivity requirement for nanoscale BPOs. Recent advances in high-sensitivity flow cytometry have made it possible to conduct precise, sensitive, and specific analyses of nanoscale BPOs, with exceptional benefits for bacteria, mitochondria, viruses, and extracellular vesicles (EVs).

Light-Scattering Sizing of Single Submicron Particles by High-Sensitivity Flow Cytometry.

Rapid and reliable size measurement of single submicron particles (100-1000 nm) is important for quality control of particulate matter, biomedical research, environmental study, and drug delivery system development. Though direct measurement of the elastically scattered light from individual submicron particles represents the simplest method for particle size measurement, the inadequate instrument sensitivity and complicated relationship between scattering intensity and particle size render it a great challenge. Combining the superior sensitivity of a laboratory-built high-sensitivity flow cytometer (HSFCM) in the side scattering (SSC) detection of single nanoparticles and the great efforts in synthesizing 38 highly monodisperse silica spheres ranging from 180 to 880 nm with small size intervals, here we report the first comprehensive comparison between the experimentally measured and Mie theory calculated intensities of light scattered by single submicron particles.

Rapid quantification of live/dead lactic acid bacteria in probiotic products using high-sensitivity flow cytometry.

A laboratory-built high-sensitivity flow cytometer (HSFCM) was employed for the rapid and accurate detection of lactic acid bacteria (LAB) and their viability in probiotic products. LAB were stained with both the cell membrane-permeable SYTO 9 green-fluorescent nucleic acid stain and the red-fluorescent nucleic acid stain, propidium iodide, which penetrates only bacteria with compromised membranes. The side scatter and dual-color fluorescence signals of single bacteria were detected simultaneously by the HSFCM.

Immobilization of chlorine dioxide modified cells for uranium absorption.

There has been a trend towards the use of microorganisms to recover metals from industrial wastewater, for which various methods have been reported to be used to improve microorganism adsorption characteristics such as absorption capacity, tolerance and reusability. In present study, chlorine dioxide(ClO2), a high-efficiency, low toxicity and environment-benign disinfectant, was first reported to be used for microorganism surface modification. The chlorine dioxide modified cells demonstrated a 10.

Laboratory investigation of reducing two algae from eutrophic water treated with light-shading plus aeration.

The occurrence of harmful algal bloom in water source poses a serious water safety problem to local water supply systems. In order to ensure the raw water quality, the feasibility of reducing harmful algae by light-shading plus aeration was investigated. The batch test showed that algal biomass reduced rapidly under light-shading condition, and the reduction efficiency was further increased when light-shading was accompanied by aeration.