100cx 2 microscope

Transmission electron microscopic (TEM) images of gold nanocubes (AuNCs) were collected using a JEOL 100CX-2 microscope and the average size of the AuNC was determined using ImageJ software. DF images and SERS spectra from the human oral squamous cell carcinoma (HSC-3) and human keratinocyte (HaCaT) cells were collected using a Renishaw inVia Raman Microscope coupled with Leica DM2500M microscope. A 785 nm diode laser was used for the SERS measurements.

The anterior parts of anesthetized A. pisum heads were severed in fixation buffer (FB; 0.1 M cacodylate buffer [pH 7.2]) under a dissecting microscope using razor blades. Samples were fixed in FB containing 0.5% glutaraldehyde and 2% paraformaldehyde and embedded in LR Gold resin. Primary antisera and secondary antibodies were used at a 1:25 dilution. Sections (60 nm thick) were observed in a JEOL 100CXII microscope operated at 60 to 80 kV.

Three aliquots of 50 μM and three of 100 uM concentration of p31-43 were prepared in MilliQ water from a 3 mM mother solution of the peptide. These solutions were deposited onto copper grids (200 mesh) coated with Formvar. After 5 min the excess fluid was removed by capillarity and the samples were negatively stained with 2% uranyl acetate in water for 2 min. After removal of excess fluid, the samples were allowed to dry and visualized using a JEOL 100CX II microscope operating at 100 kV.

Fluorescent images of particles were taken using a Zeiss Axio Observer Z1 inverted microscope. The ability of spherical capsules to conform to a circle was measured using the circularity function in ImageJ software. Transmission electron microscopy was carried out using JEOL 100CX‐II microscope operating at 100.0 kV. Particles were dropped on a Cu grid 400 mesh (Electron Microscopy Sciences) and allowed to settle for 10 min. Any excess solution was wiped off and the grid was left to dry overnight before imaging.

Treated and untreated S. aureus ATCC 25923 and A. baumannii ATCC 19606 cells were observed by transmission electron microscopy (TEM). Isolated colonies were cultured in CAMH under agitation at 37 °C until the mid-log phase. Then, 30 mL was adjusted to OD₆₀₀ = 0.05 on a SpectraMax M5 (Molecular Devices) in CAMH and treated with the peptide at 0.5×, 1×, or 4× MIC. The tubes were incubated at 37 °C for 10 min, centrifuged three times at 3000× g for 10 min, and then washed with PBS. The bacteria were resuspended in PBS with 3% glutaraldehyde and incubated at 4 °C for 2 h, followed by another centrifugation step. Cells were resuspended in PBS and fixed with 3% glutaraldehyde for 2 h at 0 °C, followed by fixation with osmium tetroxide for 2 h at 4 °C. The samples were then washed and dehydrated using increasing concentrations of ethanol. After the final ethanol wash, cells were resuspended in propylene oxide and centrifuged twice. The oxide was removed, and the material was deposited on epoxy resin and stirred overnight. Ultrafine cuts were made using an ultramicrotome. The sections were analyzed using a JEOL 100CX-II microscope (Japan). For quantitative comparison between samples, ten images were captured in random fields at 20,000× magnification. The treated and untreated samples were compared using ANOVA. Bacterial samples were prepared in biological duplicates.