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HealthSky provides Pap staining kits for staining gynecological and non-gynecological cell smears using the technique developed by George Papanicolaou.
Pap staining involves four steps: 1. Fixation; 2. Nuclear staining; 3. Cytoplasmic staining; 4. Clearing.
The main requirements are as follows: 1. Clear nuclear structure; 2. High transparency; 3. Proper differentiation of colors.
Based on the above requirements, there are several Pap staining precautions:
(1) Light nuclear staining:
1) Hydrochloric acid differentiation time is too long or hematoxylin staining time is too short. Shorten differentiation time or prolong bluing time; add a small amount of fresh hematoxylin solution daily or prepare a new hematoxylin solution.
2) Drying of the smear before fixation. Therefore, strict adherence to wet fixation principles is required for Pap staining smear preparation.
3) Tap water's pH is slightly acidic. Use an alkaline solution.
(2) Dark nuclear staining:
1) Insufficient concentration of hydrochloric acid solution. Add a few drops of hydrochloric acid to increase the concentration appropriately.
2) Fixation with alcohol concentration higher than 95% may result in over-staining. Use 95% alcohol for fixation.
(1) If the entire smear shows light cytoplasmic staining, prolong the staining time or replace with a fresh solution.
(2) If the cytoplasm is not differentiated and appears uniformly light red, it may be due to drying of the smear before fixation or interference by a large number of cocci-like bacteria. In this case, extending the staining time may partially correct the lack of differentiation.
(3) Gray or purple cytoplasmic staining is due to prolonged hematoxylin staining or poor differentiation with hydrochloric acid. Re-staining with hematoxylin after decolorization can correct this.
(4) Due to varying specimens, the same EA staining formula may cause a bias towards blue, green, or red. Different EA solutions should be used for different specimens. Generally, EA36 and EA50 are used for gynecological specimens, while EA65 or modified EA is used for non-gynecological specimens.
(5) Another important reason for lack of cytoplasmic differentiation is the inappropriate pH value of the EA staining solution. If the staining is red, adding a little phosphotungstic acid solution can correct it; if the staining is uniformly blue or green, adding a little saturated lithium carbonate solution can correct it. For modified EA staining solutions, adding 2ml of glacial acetic acid per 100ml of staining solution can enhance the staining effect and prolong the longevity of the solution.
