For goblet cell staining, sections were stained with Alcian Blue solution pH 2.5 (Sigma-Aldrich, St. Louis, MO, USA) following standard procedures. The number of goblet cells per crypt was quantified using the software NIS-Elements BR (Nikon Instruments Europe B.V.). Three sections for each animal, and 3020 crypts/section, were analyzed. To be included in the quantification, crypts had to be open toward the lumen and stretch at least two-thirds of the distance between the lumen and muscularis mucosae.
Alcian blue solution ph 2
Manufactured by Merck Group
Sourced in United States
Alcian Blue solution pH 2.5 is a laboratory reagent used for the histochemical staining of acidic polysaccharides, such as glycosaminoglycans. It provides a blue stain that allows for the visualization and identification of these components in biological samples.
Automatically generated - may contain errors
3 protocols using alcian blue solution ph 2
1
Quantifying Goblet Cell Distribution
2
Alcian Blue Staining of P. aeruginosa-Infected EVPL
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EVPL tissue pieces infected with P. aeruginosa, and uninfected tissue negative controls, were fixed, paraffin-embedded, de-paraffinized and re-hydrated as described for H & E staining. Slides were stained in 1% Alcian blue solution (pH 2.5) (Sigma-Aldrich) then rinsed using distilled water. The samples were counterstained in nuclear fast red solution (Alfa Aesar) and rinsed briefly in distilled water. Tissue sections were dehydrated in increasing ethanol concentrations and finally placed in xylene, as performed for H & E staining. Samples were mounted with DPX mounting fluid and images were taken of stained tissue sections using a Zeiss Axio Scope. A1 light microscope with the Zeiss Axiocam Erc 5s and Zeiss Zen 2.3 pro software.
3
Quantifying Goblet Cell Differentiation
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Cell differentiation was measured by goblet cell count, which were identified with alcian blue stain [34 (link)]. After deparaffinization and rehydration, samples were stained with alcian blue solution (pH 2.5) (Sigma-Aldrich, St. Louis, MO) for 5 min, then counter-stained with nuclear fast red solution (Sigma-Aldrich) for 5 min. Samples were dehydrated and mounted. The numbers and locations of alcian blue stained cells were recorded. To calculate differentiation index (DI), the number of stained goblet cells was divided by the number of total cells per crypt, then multiplied by 100.
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