Procedure 4 – 1 Quick Diff Staining Procedure
Semester Spring 2010
The Class Section MED 215-M08
Dr. Victor Veloz
The procedure that I am going to introduce is Quick Diff Staining Procedure. It is used for the differential count. After distinguishing 100 leukocytes, each of the five leukocytes are regarded as a percentage of the total 100 cells identified.
Definition of the procedure
Quick Diff Staining Procedure differentiates the distribution of the erythrocytes and five types of leukocytes on the basis of their characteristics, shapes, and sizes.
* Blood smear (On the slide with having well-feathered edge.)
* Alcohol Acetone (Fixative solution)
* Eosin solution (fluorescent red dye)
* New methylene blue (Organic staining agent)
* Staining rack (To place solutions)
* Bottled or running water (To rinse excessive solutions)
* Absorbent paper (To remove or absorb excessive solutions)
Steps of the procedure
1. Dip the slide into the alcohol acetone 3-5 times as the speed of second and remove the excessive solution with a paper towel.
2. Dip the slide into Eosin solution for 7 times and remove the excessive solution with a paper towel.
3. Dip the slide into new methylene blue for 9 times and remove the excessive solution with a paper towel.
4. Use the incinerator to allow it to dry completely.
* Erythrocytes: Pink or yellowish red.
* Platelets: Violet or purple granules.
* Neutrophils: Blue nucleus, pink cytoplasm, violet granules.
* Eosinophils: Blue nucleus, blue cytoplasm, red granules.
* Basophils: Purple or dark blue nucleus, violet granules.
* Monocytes: Violet nucleus, light blue cytoplasm.
* Bacteria: Blue.
Reference range of WBC
* Segmented neutrophil: 50 – 70 %
* Band neutrophil: 0 – 5%
* Lymphocyte: 20 – 40%
* Monocyte: 3 – 11%
* Eosinophil: 0 – 5%
* Basophil: 0 – 1%