70% Ethanol-1.5 minutes. Drain off excess. 70% Ethanol-1.5 minutes. Drain off excess. Trichrome stain-13 minutes. Drain off excess. 90% Acid Ethanol-1 3 seconds. Drain off excess. 100% Ethanol-5 10 seconds. Drain off excess. 100% Ethanol-1 minutes. Drain off excess. 100% Ethanol-1 minutes. Drain off excess. PRO-Clear-3 minutes. Drain off excess. Mount Cover Slip and examine under Oil Immersion Objective.

Permanent smears can be prepared from Unconcentrated specimen fixed in Proto-fix A specimen in Proto-fix hat has been concentrated using CONSED concentration procedure. Preparation of smear: Transfer 1-2 drops of the Proto-fix fixed specimen to a slide. May use coated slides like CELLBOND for better adhesion. Lay or hold the slide flat with specimen side up. Gently and evenly spread the sample. Spread the sample out to create thick and thin areas using a chopping motion. Allow to stand for 1-2 minutes. Or until it is dried.

Nuclear Chromatin, Chromatoid Bodies, Ingested RBCs, Bacteria- Purple to red-violet Cytoplasm of trophozoites and cysts- Blue tinted with Purple Macrophages, WBCs, Yeast- Variable- Green to blue to purple or red Background Material and artifacts blue-green to purple