During culture, metabolic activity was measured using a resazurin assay (Sigma Aldrich) at day 1,7,14, 28. After 28 days, matrix formation was quantified by biochemical assessment of GAG (dimethylmethylene Blue (DMMB),SigmaAldrich) per DNA (Quant-iT-Picogreen-dsDNA-kit, Invitrogen) according to manufacture protocols. Additionally, samples were embedded in MMA, polymerized, and saw, or paraffin embedded and cut, to visualize the cell distribution (haematoxylin staining (Sigma Aldrich)) and matrix distribution, respectively. Matrix distribution was visualized with a safranin O (Sigma Aldrich), combined with fast green to stain fibrous tissue (Sigma Aldrich). Immunohistochemistry was performed to visualize type II collagen (II-II6B3, DSHB).
Haematoxylin staining
Manufactured by Merck Group
Haematoxylin is a natural dye used in histology and cytology for staining cell nuclei blue. It is a key component in various staining techniques, including Hematoxylin and Eosin (H&E) staining, which is widely used in the analysis of tissue samples.
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2 protocols using haematoxylin staining
1
Evaluating Chondrocyte Matrix Formation
2
Evaluating Chondrocyte Matrix Formation
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During culture, metabolic activity was measured using a resazurin assay (Sigma Aldrich) at day 1,7,14, 28. After 28 days, matrix formation was quantified by biochemical assessment of GAG (dimethylmethylene Blue (DMMB),SigmaAldrich) per DNA (Quant-iT-Picogreen-dsDNA-kit, Invitrogen) according to manufacture protocols. Additionally, samples were embedded in MMA, polymerized, and saw, or paraffin embedded and cut, to visualize the cell distribution (haematoxylin staining (Sigma Aldrich)) and matrix distribution, respectively. Matrix distribution was visualized with a safranin O (Sigma Aldrich), combined with fast green to stain fibrous tissue (Sigma Aldrich). Immunohistochemistry was performed to visualize type II collagen (II-II6B3, DSHB).
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