Giemsa

Purpose:

• The Giemsa stain is used for the identification of parasites . This is a common stain in hematology
• This stain is also called a Wright Giemsa (CDC – DPDx – Diagnostic Procedures – Blood Specimens)
• The Giemsa stain can be used to identify protozoa, rickettsia, legionella, ect (Shapiro and Mandy), (Woods and Walker).

Control Tissues:

• Most often this will be an internal control to the smear itself. However, a tissue section with Giemsa positive microorganism can be created or purchased

Staining Pattern:

• Protozoa and Microorganisms – Dark Blue
• Nuclei- Blue
• Background- Pink to Pale Blue

Biochemistry Theory:

• This staining class is composed of two active dyes; Azure B and Eosin Y. When combined in the presence of particular molecular structures they yield a purple color. This process is called the Romanowsky effect, this effect can not occur with either agent by itself (Wittekind).

Troubleshooting:

• The importance of fixation is an area of debate. Some sources will state that the form of fixation is not critical (Suvarna et al.), while others insist that the fixation is important and staining quality is better in alcohol based fixatives (Barcia). Care should be taken in the lab when validating to ensure that all applicable fixation types yield consistent staining

Additional References:

• Barcia, Juan José. “The Giemsa Stain: Its History and Applications.” International Journal of Surgical Pathology, vol. 15, no. 3, 2007, pp. 292–96, https://doi.org/10.1177/1066896907302239.
• CDC – DPDx – Diagnostic Procedures – Blood Specimens. 8 Jan. 2019, https://www.cdc.gov/dpdx/diagnosticprocedures/blood/staining.html.
• Shapiro, Howard M., and Francis Mandy. “Cytometry in Malaria: Moving beyond Giemsa.” Cytometry Part A, vol. 71A, no. 9, 2007, pp. 643–45, https://doi.org/10.1002/cyto.a.20453.
• Suvarna, S. Kim, et al., editors. Bancroft’s Theory and Practice of Histological Techniques. Eighth edition, Elsevier, 2019.
• Wittekind, D. H. “On the Nature of Romanowsky-Giemasa Staining and Its Significance for Cytochemistry and Histochemistry: An Overall View.” The Histochemical Journal, vol. 15, no. 10, 1983, pp. 1029–47, https://doi.org/10.1007/BF01002498.
• Woods, G. L., and D. H. Walker. “Detection of Infection or Infectious Agents by Use of Cytologic and Histologic Stains.” Clinical Microbiology Reviews, vol. 9, no. 3, 1996, pp. 382–404, https://doi.org/10.1128/CMR.9.3.382.