In addition, both cells and matrix were stained throughout the fibrotic regions of the synovium.
Highly sulphated mucins, characteristic for cartilage-like matrix, were stained with alcian blue 8GX (in 0.5 M HCl, pH 1.5) (Sigma Aldrich) for 24 hours.
Briefly, the ethanol-fixed cells and matrix were stained for 1 h with 40 mM Alizarin red-S (pH 4.2) and extensively rinsed with water.
Briefly, ethanol-fixed cells and matrix were stained for 1 h with 40 mM alizarin red-S (pH 4.2) and rinsed extensively with water.
Thus, during a general H&E staining experiment, the nuclei of the cells are stained blue, whereas the cytoplasm and extracellular matrix are stained varying degrees of pink [ 57].
Endothelial cells were immunostained with VE-Cadherin (green); smooth muscle cells were stained with SM22a (cyan), and extracellular matrix was stained with collagen type IV (red).
The pericellular matrix was stained by using the rat anti-heparan sulfate proteoglycan antibody (Millipore, Watford, UK) followed by the Alexa-Fluor-488 labeled anti-rat IgG (Life Technologies, Paisley, UK).
TEM analysis showed that the bacterial cells were surrounded by electron-dense fibrous matrices that were stained with ruthenium red (Figure 5).
Mineralized matrix deposition were stained by alizarin red S, which showed that the BM-MSCsFluc+/GFP+ were able to differentiate into osteogenic cells.22 Bioluminescence imaging (BLI) quantification demonstrated a linear correlation between the number of BM-MSCsFluc+/GFP+ and Fluc average radiance (Figure 3c) in vitro.
At 14 days after infection, mineralized matrix nodules were stained for calcium precipitation by means of Alizarin Red S staining, as described previously [24], [25], [27], [28], [29], [30], [31], [55], [61].
