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Next, plate was washed 3 times and each well was incubated with 100 μL of TMB in the dark for 15 min at room temperature followed by 100-μL of acidic stop solution
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Fluorescence microscopy was performed at room temperature using a Nikon ECLIPSE Ni microscope equipped with a Plan Apo Fluor 60×oil immersion objective (NA 1.4) and a Clara CCD camera (Andor Technology)
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