Contact Mode Atomic Force Microscopy as a Rapid Technique for Morphological Observation and Bacterial Cell Damage Analysis.

Electron microscopy is one of the tools required to characterize cellular structures. However, the procedure is complicated and expensive due to the sample preparation for observation. Atomic force microscopy (AFM) is a very useful characterization technique due to its high resolution in three dimensions and because of the absence of any requirement for vacuum and sample conductivity. AFM can image a wide variety of samples with different topographies and different types of materials. AFM provides high-resolution 3D topography information from the angstrom level to the micron scale. Unlike traditional microscopy, AFM uses a probe to generate an image of the surface topography of a sample. In this protocol, the use of this type of microscopy is suggested for the morphological and cell damage characterization of bacteria fixed on a support. Strains of Staphylococcus aureus (ATCC 25923), Escherichia coli (ATCC 25922), and Pseudomonas hunanensis (isolated from garlic bulb samples) were used. In this work, bacterial cells were grown in specific culture media. To observe cell damage, Staphylococcus aureus and Escherichia coli were incubated with different concentrations of nanoparticles (NPs). A drop of bacterial suspension was fixed on a glass support, and images were taken with AFM at different scales. The images obtained showed the morphological characteristics of the bacteria. Further, employing AFM, it was possible to observe the damage to the cellular structure caused by the effect of NPs. Based on the images obtained, contact AFM can be used to characterize the morphology of bacterial cells fixed on a support. AFM is also a suitable tool for the investigation of the effects of NPs on bacteria. Compared to electron microscopy, AFM is an inexpensive and easy-to-use technique.

Two-Step Triethylamine-Based Synthesis of MgO Nanoparticles and Their Antibacterial Effect against Pathogenic Bacteria.

Magnesium oxide nanoparticles (MgO NPs) were obtained by the calcination of precursor microparticles (PM) synthesized by a novel triethylamine-based precipitation method. Scanning electron microscopy (SEM) revealed a mean size of 120 nm for the MgO NPs. The results of the characterizations for MgO NPs support the suggestion that our material has the capacity to attack, and have an antibacterial effect against, Gram-negative and Gram-positive bacteria strains. The ability of the MgO NPs to produce reactive oxygen species (ROS), such as superoxide anion radicals (O2•-) or hydrogen peroxide (H2O2), was demonstrated by the corresponding quantitative assays. The MgO antibacterial activity was evaluated against Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli bacteria, with minimum inhibitory concentrations (MICs) of 250 and 500 ppm on the microdilution assays, respectively. Structural changes in the bacteria, such as membrane collapse; surface changes, such as vesicular formation; and changes in the longitudinal and horizontal sizes, as well as the circumference, were observed using atomic force microscopy (AFM). The lipidic peroxidation of the bacterial membranes was quantified, and finally, a bactericidal mechanism for the MgO NPs was also proposed.

One-pot green synthesis of gold and silver nanoparticles using Rosa canina L. extract.

This study reports a green, simple, and fast method for the synthesis of gold and silver nanoparticles using natural antioxidant compounds. The aqueous extract from dried rosehips (pseudofruit of Rosa canina L.) was used as a reducing and capping agent of HAuCl4 and AgNO3 during the noble metal colloid synthesis at room temperature and no other chemical reagent was used. The high antioxidant activity of the plant extract was proven by 2,2-diphenyl-1-picrylhydrazyl assay by a spectrophotometric method. The formation of stable gold and silver nanoparticles was observed by UV-visible spectroscopy and the evolution of their characteristic surface plasmon resonance band was followed over several days. Transmission electron microscopy confirmed the formation of quasi-spherical nanoparticles with mean diameters 26 and 34 nm, for gold and silver nanoparticles, respectively; XRD revealed an FCC crystalline structure for both gold and silver NPs. The effects of concentrations of noble metal precursor and plant extract solution on the formation, stabilization and size of nanoparticles are discussed, as well as some applications of these colloids.