hi
I am PhD student in my research protocol i need to count non squamous cell in the sputum.
I'm currently using the May Grunwald's Giemsa ( MGG) but eosinophil granule not stained pink..
anybody have any idea about the staining procedure please inform me 🙏🙏
Which buffer do you use to dilute Giemsa and to wash the slides?
Note that the eosinophilic granules stain yellow/orange, not pink
I agree with antonio. The smears are satisfactory. All Romanowsky stains are very sensitive to pH of diluting buffer. After washing also you have to keep the slides in the same buffer for some time, which can be adjusted as per your satisfaction by trial and error. Further these stains show variation from different manufacturers and some times in different batches. When in hematology I prepared these stains, I always tested for optimun pH, dilution and time of staining for blood smears every time I prepared new stain. Eosinophil grnules may not be very bright red, but they are identified by their large size and uniformity, usually not overapping the nucleus. Usually with MGG (popular in Europe), pH 6.8 is recommanded. In America Write-Giemsa is popular as single stain and pH recommanded is 6.4. A pH change of 0.1 to 0.2 units may change the shade of stain significantly.
Dear Hamdia,
you can use wright stain to overcome this issue easily.
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