Hemoglobin H will also precipitate in the presence of isopropyl alcohol. The top layer of red cells from a preparation of packed red cells is incubated with brilliant cresyl blue for one hour. After incubation the upper layer is examined for blue-green inclusions. The specimen of choice is EDTA whole blood. Heinz bodies will also stain but stain blue-black and are located at the periphery of the cell.
Reticulocytes stain a dark purple-black.
During heme production iron is inserted into protoporphyrin. If iron is deficient free protoporphyrin accumulates in the cell. Protoporphyrin can be measured directly by fluorescence. The specimen of choice is EDTA whole blood. Heparinized blood can be used.