1. Specimen- Peripheral smears should be prepared from a freshly drawn blood specimen
(EDTA purple top tubes). Allow smears to air dry completely before staining.
2. Please use an X or check mark to indicate that all stains and reagents are present on the bench
before manual staining of rack. Reagents needed for manual testing are listed below.
___________ Methanol for Wright-Giemsa Stain
___________ Wright-Giemsa Stain
___________ Wright-Giemsa Stain Buffer
___________ Wright-Giemsa Stain / Buffer mixture
Please mix the Wright-Giemsa Stain and buffer mixture before blood smears are made or before the smears are air drying.
Combine 15ml of Wright-Giemsa Stain with 75ml of Wright-Giemsa Stain Buffer, mix well and allow to stand for at least 10 minutes. Make sure that a metallic sheen is observed on the surface before staining.
llow to stand 10 minutes prior to use.
WRIGHT -GIEMSA STAIN STAIN PROCE DURE
1. Place freshly prepared and air dried blood smears in a manual staining rack.______
2. Place slide rack with air dried blood smears in Methanol for 30 seconds. ______
(Adjust timer for 30 seconds.)
3. Take a disposable pipette and flood the Wright-Giemsa Stain on the appropriately labeled slides.
4. Set timer for 2 minutes. ______
5. Place oxidizing Wright-Giemsa Stain and Wright-Giemsa Stain Buffer Mixture on
Wright-Giemsa stained slides laying on slide rack. (Displacing the Wright-Giemsa Stain off the
slides with the pipette filled with Wright-Giemsa Stain/Buffer Mixture and viewing a metallic
sheen on the top of slides.) ______
6. Set timer for 6 minutes. ______
7. Place slides in Wright-Giemsa Stain Buffer for 2 minutes.
8. Set timer for 2 minutes._________
9. Retrieve slide rack with stained slides from buffer rinse and allow to air dry before examination
with immersion oil and 100X objective.
STAINE STAINED SLIDE TI PS
1. Wipe excess stain from back of slides with methanol soaked gauze, being careful not to wipe
off the stained side of the slides.
2. Place on slide envelope or slide flat/folder and proceed to the microscope room.
3. Place 1 drop of immersion oil onto slide to be viewed making sure it is completely air dried
B. Staining characteristics
Erythrocytes light pink to moderate purple, not grey or blue
Polymorphonuclear Neutrophils blue to dark blue to purple nuclei, reddish purple lilac granules,
pale pink cytoplasm
Eosinophils blue to dark blue to purple nuclei, red to orange-red granules, blue cytoplasm
Basophils purple to dark blue to black nuclei, purple granules
Lymphocytes and monocytes dark purple nuclei, sky blue cytoplasm
Platelets violet to purple granules