Three antibody staining protocol for immunolabeling of mouse brain sections
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Description
Quantitative imaging and analysis of fluorescently stained tissue sections is sensitive to multiple factors. Two critical ones are endogenous autofluorescence and careful preparation of antibodies. This protocol provides methods for reducing the impact of autofluorescence through high intensity broad wavelength light exposure. Additionally, it outlines a stepwise protocol for creating primary antibody Master Mixes which are then subsequently combined together, or with buffer, to ensure consistency in antibody concentrations across all conditions.
