Quemesa ccd camera

TEM was conducted as previously described (Konzack et al., 2015 (link)). The primary cortical astrocytes were fixed in 1% glutaraldehyde and 4% formaldehyde mixture in 0.1 m phosphate buffer for 10 min. The cells were detached, and fixation was continued for 1 h. After fixation, the cells were centrifuged, immersed in 2% agarose in distilled water, postfixed in 1% osmium tetroxide, dehydrated in acetone, and embedded in Epon LX 112 (Ladd Research Industries). Thin sections were cut with a Leica Ultracut UCT ultramicrotome, stained in uranyl acetate and lead citrate, and examined in a Tecnai G2 Spirit TEM (FEI Europe). Images were captured by using a Quemesa CCD camera (Olympus Soft Imaging Solutions GmbH) and analyzed with a Tecnai G2 Spirit 120 kV TEM with Veleta and Quemesa CCD cameras and a Philips CM100 equipped with CCD camera 23.

The morphological characteristics of the fabricated nanoparticles were analyzed with a Tecnai G2 Spirit transmission electron microscope (FEI Europe, Eindhoven, The Netherlands). The samples were prepared by first diluting each cellulose nanoparticle suspension with deionized water. A 7 µL droplet of 0.01 M polylysine suspension was added to the carbon-coated copper grid and after that a 7 µL droplet of diluted nanoparticle suspension was dosed on the copper grid and any excess sample was removed by touching the droplet gently with the corner of a filter paper. The samples were negatively stained with a droplet of uranyl acetate (2% w/v) placed on top of each sample and removing the excess uranyl acetate with a filter paper. After drying the grids under room temperature, they were analyzed at 100 kV under standard conditions. Images were captured with a Quemesa CCD camera and iTEM image analysis software (CE, Olympus Soft Imaging Solutions GmBH, Munster, Germany) was used to measure the average widths and lengths of the nanoparticles. The final results were averaged and standard deviations were calculated.

Tissues were fixed in 1% glutaraldehyde and 4% formaldehyde in 0.1 M phosphate buffer, pH 7.4, post-fixed in 1% OsO4, dehydrated in acetone and embedded in Epon LX 112 (Ladd Research Indus- tries, VT, USA) at Biocenter Oulu Tissue Imaging Center or alternatively fixed in 2% PFA + 2.5% glutaraldehyde in 0.1 M cacodylate buffer, pH 7.4 and post-fixed in 1% OsO4 in 0.1 M cacodylate buffer + 0.2% potassium ferricyanide, following embedding in Spurr's Low Viscosity resin at Integrated Microscopy Core at Baylor College of Medicine. Semi-thin sections (1 μm) were cut and stained with toluidine blue for light microscopic inspection and selection of regions of interest. Thereafter, thin sections (80 nm) were cut and post-stained in uranyl acetate and lead citrate. Specimens were examined using the Tecnai GS Spirit microscope (FEI Europe, Edinhoven, Netherlands) and images were acquired with a Quemesa CCD camera controlled by the iTEM software (Olympus Soft Imaging Solutions GmbH, Munster, Germany) at Biocenter Oulu Tissue Imaging Center.

One side of a nitrocellulose, carbon-coated PELCO^® Cu grid (Ted Pella Inc., Redding, CA, USA) was silanized by applying a 20 mkL droplet of a freshly prepared, 1% water solution of (3-aminopropyl)-trimethoxysilane (APTMS) for 2 min, followed by triple washing in water. The grid was placed into an as-synthesized AgNP colloid for 10 min for particle adsorption due to electrostatic forces between negatively charged particles and positively charged amino groups of the APTMS coating. After that, the grid was washed three times with water and dried. The control sample contained no nanoparticles, only surface silanization. Grids were examined at 80 kV with a JEM-1400 transmission electron microscope (JEOL, Tokyo, Japan) equipped with a Quemesa CCD camera (Olympus Soft Imaging Solutions, Münster, Germany). The images were manually processed in ImageJ 1.51n (National Institutes of Health, Bethesda, MD, USA) by measuring the largest and smallest dimensions for each individual particle.

The embryos were fixed with 1% glutaraldehyde and 4% formaldehyde in 0.1 M phosphate buffer (pH 7.4) for 20 min and embedded in low‐melt agarose. Agarose‐embedded embryos were then prepared and imaged at the Biocenter Electron Microscopy Core Facility (Oulu, Finland). The embryos were postfixed with 1% osmium tetroxide (Electron Microscopy Sciences, Hatfield, PA), dehydrated in acetone, and embedded in Epon LX112 (#21210; Ladd Research Industries Inc., Williston, VT). Hydroxypropyl methacrylate (Sigma, St. Louis, MO) was used in the embedding of β‐galactosidase‐stained embryos to stabilize the X‐gal reaction product (Masahira et al., 2005 (link)). Thin and semi‐thin sections were cut throughout the embryo using a Leica Ultracut UCT microtome (Leica, Wetzlar, Germany), and toluidine blue‐stained semi‐thin sections were used to select thin sections. The thin sections were stained with uranyl acetate and lead citrate and examined using a Tecnai G2 Spirit 120 kV transmission electron microscope (FEI, Eindhoven, Netherlands). Images were captured using a Quemesa CCD camera (Olympus Soft Imaging Solutions GMBH, Münster, Germany).