The Papanicolaou method is commonly used for routine cytological staining. Papanicolaou Stain was initially used only to assess estrogen levels in vaginal epithelium and detect pathogens such as Candida and Trichomonas in the genital tract. The combination of Orange G6 with EA36 or EA50 can stain the cytoplasm in distinct bright green, blue, and pink colors. Currently, most laboratories use ready‑to‑use staining solutions; therefore, the optimal conditions for each modified staining solution should be noted. The final cytoplasmic staining should be transparent and visible, and nuclear chromatin should be easily distinguishable. The currently modified Papanicolaou staining solution contains various ions and exhibits polychromatic staining efficiency, resulting in bright and transparent cytoplasm, as well as clear structures of the nuclear membrane, nucleolus, and chromatin. The main nuclear stain is Harris Hematoxylin Stain, and the main cytoplasmic stains are EA36 Stain and EA50 Stain. Papanicolaou Stain is used for exfoliated cell specimens: the cell nuclei appear blue or black, and the cytoplasm of keratinized squamous cells appears pink or orange‑red.
Papanicolaou Stain (Papanicolaou EA65) uses EA65 for cytoplasmic staining and a non‑toxic modified hematoxylin stain for nuclear staining. EA65 is more suitable for non‑gynecological cytological smears. Papanicolaou stain is prepared from hematoxylin, orange G, eosin, phosphotungstic acid, light green, etc. It is used for processing exfoliated cell specimens such as secretions. The cell nuclei appear blue or black, and the cytoplasm of keratinized squamous cells appears pink or orange-red. This reagent is for research use only and is not intended for clinical diagnostic or other purposes.
Product Components and Storage Conditions:
P1511527
| Component
| 4×100mL
| Storage
|
P1511527A
| Hematoxylin Stain
| 100mL
| RT
|
P1511527B
| Acid Alcohol Differentiation Solution
| 100mL
| RT
|
P1511527C
| Orange G6 Stain
| 100mL
| RT
|
P1511527D
| EA65 Stain
| 100mL
| RT |
Materials Required:
1. Fixative solution (e.g., 95% ethanol-glacial acetic acid fixative), graded ethanol, bluing reagent
2. Microscope, xylene or eco-friendly dewaxing & clearing agent, 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 with distilled or tap water for 1–2 min.
6. Stain with hematoxylin stain for 3–5 min.
7. Rinse with tap water for 1–2 min.
8. Differentiate in acid alcohol differentiation solution for approximately 5–10 s.
9. Wash with water for bluing for 5–10 min, or treat with bluing reagent for 10–30 s.
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 with Orange G6 stain for 1–2 min.
14. Rinse in 95% ethanol (I) and (II) for 30–60 s each.
15. Stain with EA65 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 agent, and mount coverslip with neutral balsam.
Staining Results:
Nuclei: blue-purple or black
Cytoplasm of non-keratinized cells: pale blue or pale green
Cytoplasm of keratinized cells: pink or orange-red
Precautions:
1. This reagent has been filtered. If the staining result is unsatisfactory, it may be filtered again, and the staining solution should be replaced regularly.
2. For your safety and health, please wear a lab coat and disposable gloves during operation.
ZJ26F0434273