Giemsa Stain for Microscopy

Giemsa’s staining solution (composed of methylene blue, azure and eosin) is one of the most popular microscopic stains, commonly used in hematology, histology, cytology and bacteriology for in vitro diagnostic (IVD) use. Giemsa is classified as a versatile stain primarily due to its unique formulation. Its utility is well established in hematology for blood and bone marrow specimens, bacteriology, clinical cytology specimens, histological biopsies, and tumor samples.

Below, you’ll find how to prepare Giemsa solution for various applications, Giemsa staining protocols according to sample type and application, as well as detailed quality information regarding our Giemsa formulation.

Download our scientific poster “Giemsa: The Universal Diagnostic Stain” to view examples of Giemsa’s application in hematology, histology, cytology, bacteriology, and cytogenetics.

Our High-Quality, IVD-registered Giemsa Stain

Giemsa stain is one of the core products in our microscopy portfolio. As it is intended for the investigation of human material, our Giemsa’s solution is registered as an “in vitro diagnostic medical device” (IVD) and bears the CE mark. It is produced to the highest quality standards to enhance the visibility of cell structures and ensure clear results. Other advantages include excellent batch consistency, long shelf life, and detailed certificates of analysis (CoA).

Giemsa Staining Protocols

In Giemsa staining, it is important to carefully follow the instructions for the specific type of material being investigated in order to obtain reliable results with highly differentiated cell structures. The following procedures describe staining of blood and bone marrow smears, paraffin sections and clinical-cytological specimens.

How to prepare Giemsa’s solution for various applications:

1. Buffer solution: 
   Dissolve 1 buffer tablet in 1 L distilled water
2. Giemsa’s solution for manual staining:
   Dilute 10 mL Giemsa’s solution with 190 mL buffer solution, mix well, leave to stand for 10 min, and filter if necessary.
3. Giemsa’s solution for clinical cytology fast staining:
   Use undiluted Giemsa’s solution, stain slides with concentrated Giemsa for 1 min, followed by 2 x 1 min washes with pH 6.8 buffer solution.
4. Giemsa’s solution for detection of Helicobacter pylori:
   Use undiluted Giemsa’s solution, stain slides with concentrated Giemsa solution for 15 min, followed by 10 sec washes with each of the following solutions: acetic acid (0.1%), distilled water, 2-Propanol (3 times).
5. Giemsa’s solution for staining with staining automat: 
   Slowly add 25 mL Giemsa’s solution to 275 mL buffer solution, mix and leave to stand for 10 min, and filter if necessary.
6. Giemsa’s solution from dry dye: Dissolve 0.76 g Giemsa’s azure eosin methylene blue (dry dye) in 50 mL glycerol (85%) and heat for 1 h at 55°C on a water bath, add 50 mL methanol, leave to stand for 24 hours and filter.

Giemsa Procedure of Air-Dried Smears

Giemsa Staining of Blood Samples

For hematology, carefully dilute Giemsa’s solution in the ratio 1:20 with phosphate buffer solution acc. to Weise pH 6.4, 6.8 or 7.2. The table below shows the results for phosphate buffer solution pH 6.8.

Results

Neutrophilic Granulocytes Color Nuclei Red to violet Lymphocytes Plasma blue Monocytes Plasma grey-blue Eutrophilic granulocytes Granules light violet Eosinophilic granulocytes Granules reddish to red-brown Basophilic granulocytes Granules dark-violet Thrombocytes Violet Erythrocytes Reddish Blood parasites Nuclei bright red

Blood smear, Giemsa stain  Blood smear, Giemsa stain Blood smear, Giemsa stain Blood smear, Giemsa stain Trypanosoma blood smear, Giemsa stain Malarial Gametocyte blood smear, Giemsa stain back to top

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Pappenheim Staining of Blood Samples

Alternatively, blood samples can be stained via the Pappenheim method using a combination of May-Grünwald’s solution and Giemsa’s solution.

Pappenheim stain blood smear	Pappenheim	Pappenheim stain, blood smear	Pappenheim stain, blood smear

Giemsa Staining of Paraffin Sections

Bone Marrow Samples

For bone marrow samples, the specimen should be decalcified in a gentle decalcifying solution, such as OSTEOSOFT®, so that immunohistochemistry and molecular pathology and enzymatic reactions can be successfully processed. It is also critical to use separate xylene or Neo-Clear® rinse baths, as any ethanol traces in the solutions may result in the preparations being discolored.

Pretreatment of Bone Marrow and Iliac Crest Biopsy Material

Place the fixed biopsy materials in OSTEOSOFT® decalcifying solution for 18-24 hours to gently remove any calcification, then transfer to histoprocessing. Carefully cut smaller blocks and, if required, treat again with OSTEOSOFT® for an additional 20 minutes.

Staining Procedure Step Time Deparaffinate and rehydrate the sections using standard methods Distilled water 10 sec. Undiluted, filtered Giemsa's solution 15 min 0.1% acetic acid 10 sec. Distilled water 10 sec. 2-propanol 10 sec. 2-propanol 10 sec. 2-propanol 10 sec. Xylene or Neo-Clear® 5 min Use Entellan® New to cover the preparations moistened with xylene, or use Neo-Mount® to cover those moistened with Neo-Clear®

Results Cell Type Color Cell nuclei, cells Blue, dark blue Collagen, osteoid Pale blue Eosinophilic grains Red Acidophilic mucopolysaccharides, mastocytes, cartilage matrix Reddish violet Acidophilic materials Orange red Helicobacter pylori  Blue to dark-blue      Bone marrow section Lymph node section Helicobacter pylori  back to top

Giemsa Staining of Spirochaetes

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Ordering Information

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