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JoVE Journal Cancer Research
Optimization, Design and Avoiding Pitfalls in Manual Multiplex Fluorescent Immunohistochemistry
 

Optimization, Design and Avoiding Pitfalls in Manual Multiplex Fluorescent Immunohistochemistry

Article DOI: 10.3791/59915-v • 09:15 min • July 26th, 2019
July 26th, 2019 •
1, 1, 1, 1, 1, 2, 3, 2, 1, 1, 1
1Department of Surgery, University of Michigan, 2Department of Molecular and Cellular Physiology, University of Michigan, 3Department of Pathology, University of Michigan

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Multiplex fluorescent immunohistochemistry is an emerging technology that enables the visualization of multiple cell types within intact formalin-fixed, paraffin embedded (FFPE) tissue. Presented are guidelines for ensuring a successful 7-color multiplex with instructions for optimizing antibodies and reagents, preparing slides, design and tips for avoiding common problems.

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Keywords: Multiplex Fluorescent Immunohistochemistry Cell Identification Tumor Microenvironment Deparaffinizing Rehydrating Antigen Retrieval Primary Antibody Fluorophore DAPI
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