Tecnai g2 20 tem

The size distribution and polydispersity of the silver nanoparticles were determined by dynamic light scattering (DLS) using a HPPS 5001 grain size analyzer (Malvern Instruments Ltd., Malvern, UK). Transmission electron microscopy (TEM) micrographs were obtained using a Tecnai G220 TEM (FEI Company, Hillsboro, OR, USA) operated at a 300-kV accelerating voltage. TEM samples were prepared by evaporating a drop of nanoparticle solution onto a 200-mesh copper grid, which was coated with a carbon support film. UV-visible (UV-vis) absorption spectra were recorded using an UV-3010 spectrophotometer (Shimadzu Ltd, Japan). K/S absorption spectra of treated silk fabrics were tested under a D65 illuminant at 10° observer using an Ultrascan XE spectrophotometer (HunterLab Co. Ltd., Reston, VA, USA). The X-ray diffraction (XRD) patterns of the silver nanoparticles were taken in the 2θ range of 20° to 80° at a scanning rate of 2°/min using Cu Kα radiation with a model D/max3c X-ray detector diffraction system (Rigaku Ltd, Japan).
For Fourier transform infrared (FTIR) analysis, the colloidal silver solution was poured into acetone and the resulting precipitates were dried for characterization. FTIR spectra were performed on a Nicolet 5700 FTIR spectrophotometer (Thermo Electron Corporation, USA).

TEM analyses were performed using different instruments under varying operating conditions. Measurements performed at 80 kV were conducted using a JEOL JEM-2200MCO FEGTE. Samples were loaded in an N₂-filled glovebox onto a custom-made air-tight holder to minimise air exposure during transfer. Analyses performed at 200 kV were made using either a JEOL JEM-2000FX II TEM or an FEI Tecnai G² 20 TEM. TEM samples were prepared by depositing the Li₃N dry onto a 3-mm holey carbon film copper grid in an N₂-filled glovebox. Each grid was placed within a sealed container and transferred to the instrument under a stream of N₂. In both cases, a small condenser aperture was used to reduce beam damage (the result of which is evident in Supplementary Fig. 12) and evaporation due to the instability of nanoscale Li₃N under the beam.

An FEI Tecnai G2 20 TEM with a thermionic source (LaB₆) was used to determine the morphology and the size of particles. Before taking the images, particles were centrifuged, and the pellet obtained together with a little supernatant was dropped onto a carbon film coated copper grid. Excess solution was removed carefully using Whatman filter paper no. 1. The grid was then allowed to air dry and then mounted on a single-tilt holder and imaged in bright-field.

Analyses such as the scanning electron microscopy (TESCAN MIRA3 SEM, Brno-Kohoutovice, Czech Republic), transmission electron microscopy (Tecnai G2 20 TEM, FEI, Hillsboro, OR, USA), X-ray fluorescence spectrophotometry (Phillips-JEE 4B, Worcestershire, UK), energy dispersive spectroscopy (EDS, attached to SEM), and Fourier transform infrared spectrometer FT-IR (Nicolet iS10 FT-IR Spectrometer, Thermo Scientific, MA, USA) was used to determine the specifications of the synthesised IONPs. The photocatalytic process was carried out with a UV lamp (UVP UVGL–15; 4 Watt, 230 V–50/60 Hz, Analytic, Jena, Germany).

The size and dispersion of the upconversion nanoparticles was characterized using a FEI Tecnai G2 20 TEM with a beam voltage of 200 kV. Samples were prepared by placing a droplet of a 0.2 mg mL⁻¹ nanoparticles solution in water on carbon-coated copper grid.

Kidney marrow cells of adult Tg(lysC:dsRed)^nz50Tg/ Tg(BACmmp9:Citrine-CAAX)^vi003 fish (aged three months) were FACS sorted into Mmp9⁺LysC⁺ or Mmp9^-LysC⁺ fractions and spun in a cytocentrifuge (Fisher Scientific) onto slides according to the manufacturer´s instructions and stained using Pappenheim solution (Merck).
For TEM Mmp9^HILysC⁺ or Mmp9^-LysC⁺ cells were FACS sorted from adult kidneys of Tg(lysC:CFP-NTR)^vi002/ Tg(BACmmp9:Citrine-CAAX)^vi003 fish and embedded in liquid low melting agarose (3% in PBS), fixed with 2.5% glutaraldehyde in 0.1 M cacodylate buffer containing 5 mM CaCl₂, post-fixation in 1% aqueous OsO₄ and embedded in Epon812 resin. Semi-thin sections were stained with toluidine blue and images were recorded using a 100x/N.A. 1.4 lens. Ultrathin sections were contrasted with uranyl acetate and lead citrate and imaged using a FEI Tecnai G2 20 TEM. Measurements were performed using Fiji software.
For analysis of granulated neutrophils whole larvae Tg(lysC:CFP-NTR)^vi002/ Tg(BACmmp9:Citrine-CAAX)^vi003 were PTU treated, and stained at 2 dpf with Sudan Black B (Merck) according to a published protocol^{74 (link)} and imaged on a confocal microscope.