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The fixed material was stained en bloc with 2% aqueous uranyl acetate for 2 h at 4 °C, dehydrated, and embedded in epon resin.
Nuclear material was stained with 4, 6-diamidino-2-phenylindole (DAPI; Sigma-Aldrich, USA).
Attached biofilm material was stained by addition of 250 µl of 0.5% crystal violet solution (CV) to each well of the plate for 10 min. Unbound CV stain was removed by aspiration and the plate was rinsed again in tap water until no more CV was observed to dissolve in the water.
Study material was stained immunohistochemically for polySia, NCAM and IDH1-R132H point mutation.
Formalin-fixed, paraffin-embedded material was stained with hematoxylin and eosin for histologic examination.
Nuclear material was stained with a blue fluorescent dye 4", 6 -diamidino-2-phenylindole (DAPI).
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With banding, genetic material is stained with special dyes before being examined under a fluorescent microscope.
In both cases areas rich in aggregated material were stained with Th-T giving a bright green or green yellow fluorescence against a dark background.
Microscopic slides containing 5- μm sections of the tumour material were stained with antibodies, which are commercially available.
Five μm sections from the material were stained with hematoxylin and eosin (H&E) and examined under light microscopy.
Following, materials were stained with leuco-basic fuchsin in the dark at room temperature for 2 h and treated with 1 % pectinase at room temperature for 1 h to separate the connected cells.
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