Safranin O staining | |
Reagents:
1% HCl in 70% ethanol 1% acetic acid in water Weigert’s iron haematoxylin working solution 0.1% fast green in distilled water 0.1% safranin in distilled water | |
Staining procedure (at room temperature): | Time |
Incubate with Weigert‘s haematoxylin working solution | 5 min |
Differentiate in 1%HCl/70%ethanol (under gentle shaking) | 5 s |
Rinse in running tap water | 15 s |
Incubate in 0.1% safranin O | 30 s to 1 min |
Rinse in 1% acetic acid | 15 s |
Rinse in running tap water | 15 s |
Incubate with fast green 0,1% | 5–7 min |
Rinse in 96% ethanol | 15 s |
Rinse in 100% ethanol until no colour can be removed | up to 1 min |
Incubate in xylene | 5 min |
Seal the slides in permanent mounting medium (eg, Eukitt) | |
TB staining | |
Reagents:
TB stock solution 10×: 1% TB O, 1% natriumtetraborate in water. Stock solution should be filtrated two times before use. 1× TB working solution is prepared in water. | |
Staining procedure (at room temperature): | Time |
Incubate in TB working solution (1×) | 8–30 s |
Rinse in distilled water | 15 s |
Rinse in 96% ethanol (metachromatic dye becomes visible) | 10–20 s |
Dehydrate in absolute ethanol | 5 min |
Incubate in xylene | 5 min |
Mount with permanent mounting medium (eg, Eukitt) |
TB, toluidine blue.