For electron microscopy, mice were perfused with heparinized Ringer’s solution and with freshly prepared PFA and glutaraldehyde (2.5% glutaraldehyde and 2% PFA in PBS). Until further processing, brains were postfixed in Monti-Graziadei solution (2% glutaraldehyde, 0.6% PFA, 0.03% calcium chloride in 0.06 M sodium cacodylate buffer, pH 7.35) for at least 48 hours at 4 °C. After further fixation in 1% osmium tetroxide in 0.1 M cacodylate buffer for 2 h, samples (approximately 1 mm3) were dehydrated in an ascending series of ethanol and incubated in propylene oxide followed by a 1:1 mixture of propylene oxide and araldite (Sigma-Aldrich) and subsequently embedded in araldite. Ultrathin sections were cut at approximately 80 nm and were transferred to copper grids. Sections were contrasted in a contrasting system for ultrathin sections using uranyl acetate ready-to-use solution, followed by lead citrate ready-to-use-solution (all Leica Microsystems). Images of vessels smaller than 10 μm were taken by an electron microscope (Jeol JEM 1011). After putting images of a vessel into a collage by using Inkscape 1.0.1 [RRID: SCR_014479], vesicles in the range of 30 - 200 nm were manually counted from 3 capillaries per animal and luminal membrane length was measured by using ImageJ.
Araldite
Manufactured by Merck Group
Sourced in Germany, United States, Italy
Araldite is a two-component epoxy resin system designed for use in a variety of industrial applications. It is a high-performance adhesive that provides strong and durable bonds between a wide range of materials, including metals, plastics, ceramics, and composites.
Automatically generated - may contain errors
Lab products found in correlation
Glutaraldehyde, by Merck Group (8 mentions)
Epon 812, by Merck Group (8 mentions)
Bx51 microscope, by Olympus (5 mentions)
Propylene oxide, by Merck Group (4 mentions)
Colorview 2 camera, by Olympus (4 mentions)
Lead citrate, by Merck Group (3 mentions)
Jem 1011, by JEOL (2 mentions)
Uranyl acetate ready to use solution, by Leica (2 mentions)
Lead citrate ready to use solution, by Leica (2 mentions)
Lab tek chamber slide, by Thermo Fisher Scientific (2 mentions)
Osmium tetroxide, by Merck Group (2 mentions)
Cm10 electron microscope, by Philips (2 mentions)
Methylene blue, by Merck Group (2 mentions)
H 500, by Hitachi (2 mentions)
Jem 1230, by JEOL (1 mentions)
Potassium ferrocyanide, by Merck Group (1 mentions)
Em10 electron microscope, by Zeiss (1 mentions)
Jem 1010 transmission electron microscope, by JEOL (1 mentions)
Em uc6 ultramicrotome, by Leica (1 mentions)
Phosphate buffer, by Merck Group (1 mentions)
34 protocols using araldite
1
Ultrastructural Analysis of Vessel Dynamics
2
Ultrastructural Analysis of Brain Vessels
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For electron microscopy, mice were perfused with heparinized Ringer’s solution and with freshly prepared PFA and glutaraldehyde (2.5% glutaraldehyde and 2% PFA in PBS). Until further processing, brains were postfixed in Monti-Graziadei solution (2% glutaraldehyde, 0.6% PFA, 0.03% calcium chloride in 0.06 M sodium cacodylate buffer, pH 7.35) for at least 48 h at 4 °C. After further fixation in 1% osmium tetroxide in 0.1 M cacodylate buffer for 2 h, samples (approximately 1 mm3) were dehydrated in an ascending series of ethanol and incubated in propylene oxide followed by a 1:1 mixture of propylene oxide and araldite (Sigma-Aldrich) and subsequently embedded in araldite. Ultrathin sections were cut at approximately 80 nm and were transferred to copper grids. Sections were contrasted in a contrasting system for ultrathin sections using uranyl acetate ready-to-use solution, followed by lead citrate ready-to-use solution (all Leica Microsystems). Images of vessels smaller than 10 µm were taken by an electron microscope (Jeol JEM 1011). After putting images of a vessel into a collage by using Inkscape 1.0.1 (RRID: SCR_014479 ), vesicles in the range of 30–200 nm were manually counted from three capillaries for each animal, and luminal membrane length was measured using ImageJ.
3
Histological Preparation of Insect Galls
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Galls were rinsed twice for 10 min in sodium phosphate buffer (10 mM; pH 7) and fixed overnight in 3% glutaraldehyde. Dehydration of the galls was carried out in three steps of 15 min in increasing concentrations of ethanol (30, 50 and 70%), two steps of 1 h in 90% ethanol, three steps of 1 h in 100% ethanol and, finally, two steps of 15 min in 100% acetone. Subsequently, galls were embedded in Araldite (Sigma-Aldrich) following several steps of incubation in an Araldite : acetone solution: 2 h in a 1 : 3 solution, overnight incubation in a 1 : 1 solution and three steps of 1 h in a 3 : 1 solution. Finally, the galls were oriented longitudinally in silicone molds filled with 100% Araldite and maintained at 60°C for 48 h for the polymerization of the Araldite. All steps were performed at 4°C.
4
Characterization of AuNPs Delivery
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Random lung samples, harvested at one day post-injection, were used to confirm AuNPs delivery.25 (link) Tissue specimens were fixed in 2.5% glutaraldehyde for 1 h followed by post-fixation with 1% osmium tetroxide and potassium ferrocyanide (Sigma) for 1 h at room temperature. The samples were then dehydrated in an ascending series of ethanol before they were embedded in araldite (Sigma). Ultrathin sections were cut and mounted on Formvar-coated copper grids where they were doubly stained with uranyl acetate (BDH) and lead citrate (BDH) and viewed under a JEOL transmission electron microscope (JEM-1230, Japan), the most common method for AuNPs visualization.2 (link) Elemental analysis was performed using the JEOL EDX (Energy Dispersive X-ray) microanalysis unit coupled with a JEM-1230 electron microscope.
5
Ultrastructural Analysis of Cells
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TEM was conducted as previously described [56 (link)]. Cells were seeded in 60 mm Petri dishes and then treated as described for growth experiments for 72 h. Cells were fixed in 3% glutaraldehyde (Sigma-Aldrich, Merck) solution in 0.1 M phosphate buffer (pH 7.4). Then the samples were post-fixed in osmium tetroxide (3%), dehydrated in graded acetone, and embedded in Araldite (Sigma-Aldrich, Merck). Ultrathin sections were collected on copper grids and contrasted using both lead citrate and uranyl acetate. Grids were examined in a Zeiss EM 10 electron microscope (ZEISS, Oberkochen, Germany).
6
Ultrastructural Analysis of Cells
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Cells were seeded on Lab-Tek chamber slides (Nunc), washed and fixed with 3% glutaraldehyde. Then, they were post-fixed in 1% osmium tetroxide for 1 h, rinsed, dehydrated, incubated for 2 h with 2% of uranyl acetate and embedded in Araldite (Sigma-Aldrich). Ultrathin sections were cut, stained with lead citrate, and examined under a Philips CM10 transmission electron microscope.
7
Transmission Electron Microscopy of Cells
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Transmission Electron Microscopy was conducted as previously described [9 (link)]. Briefly, cells treated as indicated were fixed in 3% glutaraldehyde (Sigma-Aldrich, Milan-Italy) solution in 0.1 M phosphate buffer (pH. 7.4) for 2h. Then the samples were post-fixed in osmium tetroxide (3%), dehydrated in graded acetone, and embedded in Araldite (Sigma-Aldrich, Milan-Italy). Ultrathin sections were collected on copper grids and contrasted using both lead citrate and uranyl acetate. The grids were examined in a “Zeiss EM 10” electron microscope.
8
Ultrastructural Analysis of Heart Tissue
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Small blocks of heart were fixed in 3 % glutaraldehyde (Acros Organics, Thermo Fisher Scientific, Waltham, MA, USA) in Sorensen phosphate buffer (0.1 M; pH 7.4) overnight at 4 °C, post-fixed with 1 % osmium tetroxide in 0.1 M Sorensen phosphate buffer for 30 min, dehydrated, and embedded in Araldite (Fluka-Sigma Aldrich). Semi-thin sections, 2-μm thick, were stained with toluidine blue. For electron microscopy, ultrathin sections (200 nm) were obtained with an Ultracut ultramicrotome (Reichert-Jung, Leica, Microsystems GmbH, Wetzlar, Germany), stained with uranyl acetate and lead citrate, and observed with a JEM 1010 transmission electron microscope (Jeol, Tokyo, Japan).
9
TEM Specimen Preparation and Imaging
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For transmission electron microscopy (TEM) specimens were fixed with 2.5% glutaraldehyde and post-fixed with 1% OsO4. Specimens were embedded in Araldite (Sigma-Aldrich). Ultrathin sections (60–80 nm) were cut with an Ultramicrotome PowerTome XL, equipped with a Drukkert 45° diamond knife, and contrasted with uranyl acetate. Ultrathin sections were studied under a Zeiss-1000 (Carl Zeiss) transmission electron microscope. The images were processed using ImageJ software (FiJi).
10
Morphological Changes of THP-1 Cells under Lactobacilli
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Changes in the morphology of the THP-1 cells following stimulation with lactobacilli were observed under a light microscope (Nikon, Tokyo, Japan). Selected lactobacilli and THP-1 cells, treated and untreated as indicated, were sequentially fixed with 2.5% glutaraldehyde and 1% osmium tetroxide (OsO4) in 0.1 M cacodylate buffer (pH 7.4). After dehydration with graded ethanol, cells were embedded in araldite (Sigma-Aldrich, Gallen, Switzerland) and sectioned. Ultrathin sections (4 μm) were counterstained with uranyl acetate and lead citrate and examined with a Leica electron microscope (Leica, Solms, Germany).
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