Staining solution reagent Luxol fast blue Kluwer Barrera

for cytologyfor histology

Minimum number of tests that can be performed 100 Completion time 20 minutes + overnight Shelf life 2 years Storage conditions 15-25°C Additional equipment Not required Application Method indicated for showing myelin and phospholipids on histological sections. Result Myelin turquoise blue Neurons and glial nuclei from pink to violet Nissl substance pale pink Product for the preparation of cyto-histological samples for optical microscopy. To show myelin and phospholipids in histologic sections. PRINCIPLE Luxol fast blue dye is a derivative of tetrabenzotetrazo-porphyrin. Kluver has demonstrated porphyrins have a selective affinity for myelin (see references). Luxol fast blue’s affinity for central nervous system is usually ascribed to the bonds it forms with phospholipidic structures such as lecithin and sphingomyelin. METHOD 1) Deparaffinise and bring section to ethanol 95°. 2) Prepare the incubation box by adding some drops of distilled water on filter paper in Petri dish and lay down the slide; put on the slide 10 drops of reagent A, close the incubation box and incubate at 56°C overnight in oven. 3) Extract the slide from oven and wash it with ethanol 95° (crystalline residues of reagent A should melt). 4) Wash in distilled water. 5) Put on the section 10 drops of reagent B: leave to act 30 seconds. 6) Differentiate in ethanol 70° until myelinic fibres become blue on colourless background (Sometimes differentiation can be difficult; repeat the step 5 for 30 seconds and put the slide again in ethanol 70°) 7) Wash well in distilled water (at least 2 times).