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Remove air bubbles from the syringe by holding it needle-up and tapping gently, then pushing the plunger slightly to expel trapped air

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To facilitate staining development, 30 l (equivalent to 1 drop) of DAB Chromogen was added to 1.0 ml of DAB Buffer (Epredia, TL-125-QHD), agitated to achieve homogeneity, and subsequently applied to the tissue sections for 10 min