Author: Kelsey Goldman, HTL (ASCP)

Last Update: December 28 2023

Copyright: 2023,

Cite this Page! -> Goldman, Kelsey. “Giemsa.” HistologyOutlines.Com, December 28 2023. Accessed (today) 

Special Stain


  • 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).


  • 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,
  • CDC – DPDx – Diagnostic Procedures – Blood Specimens. 8 Jan. 2019,
  • Shapiro, Howard M., and Francis Mandy. “Cytometry in Malaria: Moving beyond Giemsa.” Cytometry Part A, vol. 71A, no. 9, 2007, pp. 643–45,
  • 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,
  • 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,


Brancoft's Giemsa Method

Giemsa stock solution– mix and dissolve at 60C then add methanol

  • Giemsa stain powder 8g
  • Glycerol 250 mL
  • Methanol 25 mL

Working solution

  • 4mL Giemsa stock
  • 96 mL acetate buffered deionized water 


  • Deparaffinization and dehydrate
  • Rinse in 6.8 pH buffered diH2O
  • Stain in working Giemsa over night
  • Quick wash in deionized water 
  • Rinse in 0.5% acetic acid until the section turns pink
  • wash in tap water. Blot dry and dehydrate through alcohol and mount

(Suvarna et al.)

Stock Solution- a 100x Giemsa buffer

  • 54.24g of Na2HPO4
  • 36.38g of NaH2PO4H2O
  • into 1000 mL of diH2O

Working Giemsa buffer-  Make Fresh 

  • 10 mL of the stock buffer into 990 mL diH2O- the pH of this solution should be 7.2
  • Triton X 5%

Stock Giemsa solution can now can be purchased. ensure that it is stored in a cool dry place


  • Place the above into a copin jar with the slides for 45-60 minutes
  • rinse 3-4 times in Giemsa buffer
  • air dry
  • smears can be fixed in methanol 100%

(CDC – DPDx – Diagnostic Procedures – Blood Specimens)