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PAH & E Staining – Step by Step Procedure
1️⃣ Deparaffinization
Slide को Xylene I – 5 min
Xylene II – 5 min
➡️ इससे paraffin wax हट जाता है।
2️⃣ Hydration (Alcohol series)
Absolute alcohol – 2 min
95% alcohol – 2 min
70% alcohol – 2 min
Running tap water – 2 min
3️⃣ Hematoxylin staining
Slide को Hematoxylin solution में 5–10 min रखें।
➡️ Nucleus blue/purple हो जाता है।
4️⃣ Washing
Running tap water – 5 min
5️⃣ Differentiation
1% Acid alcohol – few seconds
➡️ Extra stain हटाने के लिए।
6️⃣ Bluing
Running tap water / ammonia water – 2–3 min
➡️ Nucleus clear blue हो जाता है।
7️⃣ Counterstain (Eosin)
Eosin solution – 1–2 min
➡️ Cytoplasm pink हो जाता है।
8️⃣ Dehydration
70% alcohol – few seconds
95% alcohol – few seconds
Absolute alcohol – 1 min
9️⃣ Clearing
Xylene I – 2 min
Xylene II – 2 min
🔟 Mounting
DPX mounting medium डालकर coverslip लगा दें।
✅ Result (Microscopy):
Nucleus – Blue/Purple
Cytoplasm – Pink
RBC – Bright pink/red
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#Hematoxlyin
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