Procedure for Leishman Staining and Differential WBC Count

Procedure of Leishman Staining

• Let thin blood film to air dry
• Then flood the slide with Leishman stain solution
• Wait for 8 minutes. Then flood off with distilled water in 2 – 3 seconds. Longer washing will remove stain
• Wait for 20 minutes. Observe the appearance of metallic cum that float on the surface of the slide.
• Then wash with tap water. Stand in a rack to drain and air dry.

Perform differential white blood cells count

• Using a low-power lens (10 x objectives) on the microscope, check the approximate differential cell distribution.
• Using the 40x objective high-power lens, perform a 200 cell differential count.
• 100x objective oil immersion lens should be used for examining fine intracellular details of the cells.
• Move the slide along the stage of the microscope in a broad battlement track running transversely across the body of the film, avoiding the edges completely.
• Record the numbers of each type of white cell. Calculate the percentage of each of the five basic leucocytes.