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Then the preparations were washed with 10% Phosphate Buffer Saline (PBS) 3 times for 5 minutes, dropped with Chromogen 3,3-diaminobenzidine for 510 seconds, washed with distilled water, counterstained with Hematoxylin Mayer for 510 seconds followed by washing with running tap water for 1015 minutes

Allow both to air-dry for 1015 seconds

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