<figcaption>In vivo imaging of the coexpression of endothelial adhesion molecules E-selectin and P-selectin in the mouse skin. Red, E-selectin; green, P-selectin. Credit: Charles P. Lin, Harvard Medical School</figcaption>
In vivo imaging of the coexpression of endothelial adhesion molecules E-selectin and P-selectin in the mouse skin. Red, E-selectin; green, P-selectin. Credit: Charles P. Lin, Harvard Medical School

User:

Charles Lin, Associate Professor of Dermatology, Harvard University Medical School

Project:

Investigating the role of vascular endothelial cell adhesion molecules in the entry of circulating cancer cells into tissue

Problem:

Immunofluorescent labels may bind to targets other than the specific antigen for which they are designed. In vitro protocols include a washing step to minimize the effect of nonspecific binding, but that's not possible in vivo.

Solution:

In vivo immunofluorescence protocols differ considerably from in vitro protocols, but that difference allows researchers to add a robust control for nonspecific binding, says Lin.

In vitro immunofluorescence is often performed as a sandwich-type assay,...

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