The Papanicolaou method is commonly used for routine cytological staining. Papanicolaou Stain was initially used only to detect vaginal epithelial estrogen levels and pathogens such as Candida and Trichomonas in the genital tract. The combination of Orange G6 with EA36 or EA50 allows the cytoplasm to be stained in distinct colors including green, blue, and pink. At present, most laboratories use ready‑to‑use staining solutions, so the optimal conditions for each modified staining solution should be noted. The final cytoplasmic staining should be transparent and clear, and nuclear chromatin should be easily distinguishable. Currently, modified Papanicolaou staining solutions contain various ions and exhibit polychromatic staining efficiency. After staining, the cytoplasm is bright and transparent, and the structures of the nuclear membrane, nucleolus, and chromatin are clear. The main nuclear stain is Harris hematoxylin solution, and the main cytoplasmic stains are EA36 solution and EA50 solution. Papanicolaou Stain is used for exfoliated cell specimens. Cell nuclei appear blue or black, and the cytoplasm of keratinized squamous cells appears pink or orange‑red.
Papanicolaou Stain Solution (Papanicolaou EA36) uses EA36 solution for cytoplasmic staining and non‑toxic modified hematoxylin solution for nuclear staining. EA36 is more suitable than EA50 for gynecological cytological smears, such as screening for cervical cancer and precancerous lesions. This reagent is for research use only and is not suitable for clinical diagnosis or other purposes.
Product Components and Storage Conditions:
P1511562
| Component
| 4×100mL
| 4×500mL
| Storage
|
P1511562A
| Hematoxylin Stain
| 100mL
| 500mL
| RT. Store in the dark.
|
P1511562B
| Acid Alcohol Differentiation Solution
| 100mL
| 500mL
| RT
|
P1511562C
| Orange G6 Stain
| 100mL
| 500mL
| RT. Store in the dark.
|
P1511562D
| EA36 Stain
| 100mL
| 500mL
| RT. Store in the dark. |
Materials Required:
1. Fixative solution (e.g., 95% ethanol fixative)
2. Graded ethanol series
3. Microscope
4. Xylene, neutral balsam
Protocol (For Reference Only):
1. Fix cell smears in 95% ethanol for 15 min.
2. Immerse in 95% ethanol for 30–60 s.
3. Immerse in 80% ethanol for 30–60 s.
4. Immerse in 70% ethanol for 30–60 s.
5. Rinse in distilled or tap water for 1–2 min.
6. Stain in hematoxylin stain for 3–5 min.
7. Rinse in tap water for 1–2 min.
8. Differentiate in acid alcohol differentiation solution for about 5–10 s.
9. Rinse in tap water for bluing for 5–10 min.
10. Dehydrate in 70% ethanol for 30–60 s.
11. Dehydrate in 80% ethanol for 30–60 s.
12. Dehydrate in 95% ethanol for 30–60 s.
13. Stain in Orange G6 stain for 1–2 min.
14. Rinse in 95% ethanol (I) and (II) for 30–60 s each.
15. Stain in EA36 stain for 3–5 min.
16. Dehydrate in absolute ethanol (I) and (II) for 1–2 min each.
17. Clear in xylene or dewaxing clearing solution, and mount with neutral balsam.
Staining Results:
Nuclei: bluish-purple or black
Cytoplasm of non-keratinized cells: pale blue or pale green
Cytoplasm of keratinized cells: pink or orange-red
Precautions:
1. This product has been filtered. If staining is unsatisfactory, it may be filtered again. The staining solution should be replaced regularly.
2. For your safety and health, please wear a lab coat and disposable gloves during operation.
3. This product is for research use only, strictly prohibited for other purposes.
ZJ26F0434449