Ag NPs (20 and 200 nm) were obtained from PlasmaChem GmbH (Berlin, Germany). Silver nitrate (AgNO3) was purchased from VWR (Leuven, Belgium) and Alcian blue, H2O2, Toluidine blue, 2′,7′-dichlorofluorescin diacetate (DCFH-DA), osmium tetroxide (OsO4), glutaraldehyde, resazurin sodium salt and the epoxy resin embedding kit from Sigma Aldrich (Bornem, Belgium). IL-8 kit was obtained from ENZO Life Sciences BVBA (Zandhoven, Belgium).
Alcian blue
Manufactured by Avantor
Sourced in United Kingdom
Alcian Blue is a staining dye used in histology and microscopy to detect the presence of acidic polysaccharides, such as glycosaminoglycans, in biological samples. It binds to these molecules, allowing their visualization and identification under a microscope. Alcian Blue staining is commonly used in the analysis of cartilage, mucus, and other extracellular matrix components.
Automatically generated - may contain errors
Lab products found in correlation
Toluidine blue, by Merck Group (1 mentions)
Silver nitrate agno3, by Avantor (1 mentions)
Glutaraldehyde, by Merck Group (1 mentions)
Resazurin sodium salt, by Merck Group (1 mentions)
Histoclear, by Thermo Fisher Scientific (1 mentions)
Picrosirius red, by Merck Group (1 mentions)
Hematoxylin qs, by Vector Laboratories (1 mentions)
Eosin y solution, by Merck Group (1 mentions)
Glycerol, by Avantor (1 mentions)
Cathepsin b, by Merck Group (1 mentions)
Brilliant blue g colloidal concentrate, by Merck Group (1 mentions)
5 protocols using alcian blue
1
Silver Nanoparticle Characterization
2
Alcian Blue Staining of Micromasses
3
Comprehensive Tissue Histological Analysis
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Paraffin-embedded tissue sections were stained with haematoxylin and eosin (H&E; Leica, Germany), Picrosirius Red (Merck Sigma Aldrich., Germany) for collagen, Elastin Van Gieson (Millers Elastin, TCS Biosciences, UK) for elastin, and Alcian Blue (AB; BDH Chemicals Ltd, Cellpath Ltd) stains for glycosaminoglycans (GAG). Appropriate positive controls were used to ensure histological stains were performed correctly (small intestine for PR and AB, lung for EVG). For all histological samples, three biological and technical replicates were assessed using standard H&E prior to any special stains to ensure homogeneity. All images shown are representative samples.
For H&E staining, cryosections of 3D cultures and 2D cultured cells on matrigel-coated dishes were fixed in 4% PFA in PBS 15 min, rinse with PBS followed by tap water. They were incubated with Hematoxylin QS (Vector Laboratories, UK) 1–2 min, rinsed with tap water 5 min, and incubated with Eosin Y solution (Sigma-Aldrich, UK) for 30 sec. They were dehydrated with 95% then 100% EtOH, dipped in Histoclear (ThermoFisher scientific, UK) and mounted in a non-aqueous mounting medium.
For H&E staining, cryosections of 3D cultures and 2D cultured cells on matrigel-coated dishes were fixed in 4% PFA in PBS 15 min, rinse with PBS followed by tap water. They were incubated with Hematoxylin QS (Vector Laboratories, UK) 1–2 min, rinsed with tap water 5 min, and incubated with Eosin Y solution (Sigma-Aldrich, UK) for 30 sec. They were dehydrated with 95% then 100% EtOH, dipped in Histoclear (ThermoFisher scientific, UK) and mounted in a non-aqueous mounting medium.
4
Dual Staining of Froglet Bone and Cartilage
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Staining of bone and cartilage in froglets has been previously described (Barker and Beck, 2009 (link)), and was undertaken 2 months after amputation. Briefly, froglets had been fixed and stored in 4% formaldehyde (v/v) in PBS. They were cut in half just below the forelimbs and the posterior ends discarded. The viscera were removed from the abdominal cavities and the froglets were rinsed in PBS. They were bleached using 5% H2O2 in PBS under a warm light bulb on a nutator during the day and kept stationary at night, for three days with solutions changed daily. Once the pigment had been completely bleached, froglets were rinsed several times with PBS and bubbles were removed from the eyes using fine forceps. Froglets were washed briefly in 70% EtOH in PBS and stained with Alcian Blue stain for cartilage (10 mg Alician Blue [BDH laboratory supplies] in 60 mL of EtOH + 40 mL acetic acid) for 8 hours. Tissue was washed three times with 70% EtOH (v/v) in PBS over a period of 2 hours and then macerated using 1% KOH for 5 days with daily KOH changes. They were then stained for mineralized bone using Alizarin Red S (10 mg in 100 mL of 1% KOH) for 2 h before soaking overnight in 1% KOH. Bone and cartilage stained samples were cleared and stored in a 1:1 ratio of glycerol (BDH laboratory supplies) and 95% EtOH.
5
Cathepsin-B Digestion of PGATyr
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The poly(L-glutamic acid-L-tyrosine) 4:1, sodium salt (PGATyr) (MW: 20–50 kDa; Sigma-Aldrich, UK) (2 mg/mL) was incubated with cathepsin-B (from human placenta; Sigma-Aldrich, UK) (0.6 units/mL) in pH 6.4 and pH 7.4 solutions, for 3 days, in an orbital incubator, at 37 °C. 15 μL aliquots were obtained from the incubating mixture every 24 h and were stored at −20 °C until use. Samples were treated with tris-glycine sodium dodecyl sulphate (SDS) sample buffer (Novex by Life Technologies, UK) as recommended by the manufacturer prior to gradient SDS/ polyacrylamide gradient gel electrophoresis (PAGE). After electrophoresis, gels were stained with 0.5% Brilliant Blue G-Colloidal concentrate (Sigma-Aldrich, UK) for 24 h to expose the protein ladder bands, followed by staining with 0.5% Alcian blue (BDH chemicals, UK) for visualising PGATyr. The gel bands were analyzed using ImageJ® for determining the progress of digestion within the course of 3 days.
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