STEP 2: Lake the thick smear by placing it in a small amount of water to which a few
drops of methylene blue stain has been added. Dip the slide three to five
times. Allow the slide to dry in a vertical position with the thick smear at the
STEP 3: Stain the whole slide in a 1:50 Giemsa/Triton-buffered water solution for 45
STEP 4: Rinse the thin film by dipping three times in 0.01 percent Triton-buffered water,
immerse the thick film an additional three to five minutes (a longer time may be
required for slides which have been prepared longer).
STEP 5: Allow the slide to air-dry and examine it under the oil immersion objective of
STEP 6: For the preservation of the slide, mount it using a suitable mounting medium.
Figure 2-4. The Giemsa (Triton X-100) technique for staining blood films.
(a) The nuclear portion of the malaria parasite stains magenta-red, while
the cytoplasm stains light blue. The structural details of the organism are sharper and
clearer with the Giemsa-Triton stain. The Schuffner's granules stain especially brilliant
with this stain. The undulating membrane and flagella of trypanosomes are distinctly
visible. In the microfilariae, the nuclear column is sharper and structures such as the
inner body, the mouth, and the tail structures are more visible. The sheath does not stain
well but it is visible.