Scaffolds were fixed for 4 h in a 4% paraformaldehyde solution at 4 °C and washed overnight at RT in a solution containing 100 mM sodium cacodylate and 50 mM BaCl2 at pH 7.4. The scaffolds were then dehydrated following standard procedures and embedded in a solution containing paraffin wax (Sigma-Aldrich, Madrid, Spain) and absolute ethanol (1:1), overnight at 37 °C. Next, the samples were incubated for 1 h in 100% paraffin wax and finally the blocks were made at RT in a dry atmosphere using silica gel as desiccant (Sigma-Aldrich, Madrid, Spain). Five-µm slices were obtained, deparaffinized, rehydrated and stained with hematoxylin-eosin following standard procedures. For Sirius red staining, slices were incubated with a Sirius red solution (1% Sirius Red F3BA in saturated picric acid aqueous solution) for 45 min at RT. The samples were then dehydrated and mounted with Entellan (Merck, Madrid, Spain). A Leica DM4000 microscope and digital camera DFC (Leica, Madrid, Spain) were used to analyze the samples.
Paraffin wax
Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom, Spain, India, Switzerland, Mexico
Paraffin wax is a refined petroleum-based wax used in various laboratory applications. It has a high melting point and is solid at room temperature. Paraffin wax is commonly used as a medium for embedding and mounting samples for microscopic analysis.
Automatically generated - may contain errors
Lab products found in correlation
Xylene, by Merck Group (10 mentions)
Ethanol, by Merck Group (9 mentions)
Sodium chloride (nacl), by Merck Group (5 mentions)
Toluene, by Merck Group (5 mentions)
Methanol, by Merck Group (5 mentions)
Chloroform, by Merck Group (5 mentions)
Formaldehyde, by Merck Group (4 mentions)
Hydrochloric acid (hcl), by Merck Group (4 mentions)
Formalin, by Merck Group (4 mentions)
Streptozotocin (stz), by Merck Group (4 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (3 mentions)
Acetone, by Merck Group (3 mentions)
Acetic acid, by Merck Group (3 mentions)
Diethyl ether, by Merck Group (3 mentions)
Hematoxylin, by Merck Group (3 mentions)
Ammonium hydroxide solution, by Merck Group (3 mentions)
Ethyl alcohol, by Merck Group (2 mentions)
Ascorbic acid, by Merck Group (2 mentions)
Microtome, by Leica camera (2 mentions)
Histoclear, by National Diagnostics (2 mentions)
78 protocols using paraffin wax
1
Paraffin Embedding and Histological Staining
2
Histopathological Analysis of Murine Lung Samples
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Mice were sacrificed on day 14, and lung samples were collected for histopathological analysis. Briefly, the lungs of the sacrificed mice were collected, immediately fixed with 4% paraformaldehyde (Sigma-Aldrich) for 16 h, and processed using graded alcohol and xylene (Sigma-Aldrich) before being embedded in paraffin wax (Sigma-Aldrich). Tissue sections of 5 μm were collected on microscope slides and stained using a hematoxylin and eosin (H&E) staining kit (ab245880, Abcam) and a Masson’s Trichrome Stain Kit (Polysciences Inc., Warrington, PA, USA).
3
Immunohistochemical Analysis of Splenic Tissues in Chickens
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The splenic tissues were collected from slaughtered chickens (n = 5 from each group) for immunohistochemical analyses as previously pronounced (Kalyuzhny, 2016 ). Briefly, fixation of the splenic tissues was carried out in 10% formalin solution (Cat. No. HT501128; Sigma-Aldrich, St. Louis, MO). After that, the examined tissues were dehydrated in absolute ethanol (Cat. No. 493511; Sigma-Aldrich, St. Louis, MO), cleared in xylene (Cat. No. 534056; Sigma-Aldrich, St. Louis, MO), impregnated in paraffin wax, (Cat. No. 327212; Sigma-Aldrich, St. Louis, MO), and cut at thin sections (5 mm thicknesses) using a microtome. Then, these tissue sections were deparaffinized in xylene and rehydrated through a series of decreasing alcohol solutions and finally dipped in a jar containing distilled water for 5 min and then in 3% H2O2 methanol solution (Cat. No. H1009; Sigma-Aldrich, St. Louis, MO) for 10 min to block the endogenous peroxidase activity. After that, these tissue sections were processed for staining with a primary antibody against chicken MG (Cat. No. ab35156, R&D Systems, Minneapolis, MN) and secondary HRP-labeled anti-chicken IgG H&L (Cat. No. ab205718, Abcam, Waltham, MA). Staining with DAB brown was utilized to visualize the bacterial antigen microscopically.
4
Surfactant-based Emulsion Characterization
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Toluene, methanol, paraffin wax (ASTM D 127, MP 70 °C–80 °C), light white mineral oil (density 0.84 g/mL at 25 °C), methylene blue, methyl orange, sudan blue dyes, and Whatman filter paper (grade 4, diameter 90 mm) were purchased from Sigma Aldrich, St. Louis, MO, USA. Other oils like olive oil, sunflower oil, and corn oil were purchased from a local market. Poly(dimethylsiloxane)-b-poly(ethylene oxide) (PDMS-b-PEO, Mw: 600 g/mol, PDMS:PEO 25:75) was purchased from Polysciences Inc., Northampton, UK. All the above chemicals were used without any further purification. Deionized water was obtained from Milli-Q Advantage A10 ultrapure water purification system (Millipore, Billerica, MA, USA).
5
Nanoparticle Toxicity Evaluation Protocol
6
Balsa Veneer Preparation and Characterization
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Balsa (Ochroma pyramidale) veneers with an oven-dried density of ~0.22 g cm−3 were purchased from Material AB (Sweden). Sodium chlorite (NaClO2, 80%), sodium chloride (NaCl, 99%), and dimethyl sulfoxide (DMSO, 99%) were received from Sigma Aldrich. PEDOT:PSS (Clevios PH1000, water suspension with ≈1% solid content) was purchased from Heraeus, Germany. Carbon fibers, paraffin wax, silver paste, and carbon paste were purchased from Sigma Aldrich and used as received. Blue gel (250 g) was purchased from Cefar-Complex, Sweden.
7
Evaluating Everolimus Effect on 3D Tissues
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After treatment with everolimus, the 3D tissues were fixed in 10% neutral buffered formalin (Sigma-Aldrich). The formalin-fixed 3D tissues were placed in paraffin (wax) (Sigma-Aldrich) to create formalin-fixed paraffin embedded (FFPE) blocks which then were cut using a microtome and mounted on glass microscope slides. Subsequently, the tissues were processed for staining with hematoxylin and eosin (H and E) and evaluated microscopically to assess the effect of everolimus on the 3D tissues. All images were taken at 40× magnification by the Nikon Digital Sight camera system DS-R11.
8
Histological and Molecular Analysis Protocol
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MLT, SD, phosphoric acid (H3PO4), disodium hydrogen phosphate (Na2HPO4), formaldehyde, and paraffin wax were purchased from Sigma-Aldrich (Madrid, Spain). Potassium dihydrogen phosphate (KH2PO4), potassium hydroxide (KOH), methanol (MeOH), and phosphoric acid (H3PO4) were purchased from Panreac Quimica (Barcelona, Spain). Hanks’ Balanced Salt solution (HBSS), hematoxylin and eosin were purchased from Merck S.L. (Barcelona, Spain). Double-distilled water was obtained from a Milli-Q® Gradient A10 system apparatus (Millipore Iberica S.A.U., Madrid, Spain). TRIZol reagent and RevertAid First Strand cDNA synthesis kit were purchased from Thermo Fisher Scientific (Barcelona, Spain).
9
Synthesis of Polymeric Microspheres for Adsorption
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Styrene (St) (>99%), divinylbenzene (DVB) (80%), sodium 4-vinylbenzenesulfonate (NaVBS) (>90%), ammonium persulfate (NH4)2S2O8 (APS) (>98%), 2,2′-azobis(2-methylpropionitrile) (AIBN) (>98%), ammonium hydroxide solution (NH4·OH) (28%) and basic alumina (Al2O3) (≥98%), 3-(triethoxysilyl)propionitrile (TESPN) (97%), 3-(triethoxysilyl)propyl-methacrylate (TSPM) (99%), sulfuric acid (H2SO4) (95.0–98.0%), branched polyethyleneimine (bPEI, Mn ≈ 10,000 by GPC, Mw ≈ 25,000 by LS) and paraffin wax (mp 53–58 °C) were purchased from Sigma-Aldrich (Buchs, Switzerland). N,N’-diisopropylcarbodiimide (DIC) (99%) was purchased from Acros Organics (Basel, Switzerland). St and DVB were passed through basic alumina to remove the stabilizer before usage. AIBN was purified by re-crystallization twice from methanol and stored at −20 °C before usage. Other reagents were used as received. Ultrapure water (UPW; conductivity c = 0.055 µS/cm and resistivity, ρ = 18.2 MΩ cm at 298 K) was obtained from an Arium 611 VF water purification system (Startorius stedim biotech, Aubagne, France), and it was used as the aqueous medium in all experiments.
10
Maize Leaf and Stem Histological Analysis
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For histological analysis, at flowering time, ear leaves and the third stem internodes above ground of C01 and STTM166-M plants were collected with three biological replicates from different transformation events. Selected and cleaved the middle part of these collected maize leaf and stem samples into small pieces, further placed in precooled 70% FAA solution. After the tube air pumped, the samples were fixed overnight at 4 °C in FAA solution, and further to dehydrate in graded ethanol series approach. The samples were then re-dehydrated and transparent treated, and embedded in paraffin wax (Sigma-Aldrich). Further, the samples were cleaved into 4-μm sections using a Leica RM 2265 programmable rotary microtome (Leica Microsystems). After being stained with 0.05% toluidine blue, the sections were photographed using an Olympus IX73 microscope (Olympus, Tokyo, Japan).
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