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

DOI:
10.3791/59915-v

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09:15 min

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July 26, 2019

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, , , , , , , , , ,
1Department of Surgery,University of Michigan, 2Department of Molecular and Cellular Physiology,University of Michigan, 3Department of Pathology,University of Michigan

Chapters

  • 00:04Title
  • 00:55Deparaffinization and Rehydration
  • 02:24Slide Staining
  • 05:05DAPI Application
  • 05:55Imaging of Monoplexes and Multiplex Preparation
  • 07:18Results: Monoplex and Multiplex Optimization
  • 08:39Conclusion

Summary

Automatic Translation

Automatic Translation

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.

Tags

Keywords: Multiplex Fluorescent ImmunohistochemistryCell IdentificationTumor MicroenvironmentDeparaffinizingRehydratingAntigen RetrievalPrimary AntibodyFluorophoreDAPI

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