After CO2-induced euthanasia, multiple organs (spleen, liver, and kidney) were collected and fixed in 4% paraformaldehyde for 24 hours and processed for paraffin embedding. Tissues embedded in paraffin blocks were sectioned using a microtome (Microm HM355S, Thermo Scientific) and transferred to glass slides (FisherbrandTM Superfrost plus, Pittsburgh, PA). Tissue sections were stained with H&E solution (Hematoxylin: GHS280, Sigma Aldrich; Eosin: HT1101128, Sigma Aldrich) and imaged using a Keyence microscope (BZX-100). Tissues were analyzed by observing for splenic clustering in the white pulp regions of the spleen associated with follicular development of lymphocytes [29 ] and infiltration of mononuclear cells in the kidneys and livers associated with GVHD-like pathology [30 (link), 31 (link)].
Hematoxylin ghs280
Manufactured by Merck Group
Hematoxylin: GHS280 is a laboratory reagent used in histology and cytology. It is a blue-black stain that binds to nucleic acids, enabling the visualization of cell nuclei.
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2 protocols using hematoxylin ghs280
1
Histological Analysis of Organs in GVHD
2
Histological Analysis of Tumor Metastasis
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Fixation, processing and staining of tissue sections from tumours and lungs was carried out as previously described.37 (link) Tumours and lungs dissected from NSG mice were fixed in 10% paraformaldehyde and embedded in paraffin. Tissue was sectioned using a microtome at 10 µm thickness. For H&E staining: standard procedures were followed, including deparaffinizing, hydration, staining with Hematoxylin (GHS280; SIGMA, St. Louis, MO) and counterstaining with Eosin (HT110180; SIGMA, St. Louis, MO). Lung metastases were quantified by three independent observers blinded to treatment using ImageJ.
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