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Cancer Research

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Nanyang Technological University

Optimization, Design and Avoiding Pitfalls in Manual Multiplex Fluorescent Immunohistochemistry

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9.2K Views

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09:15 min

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July 26th, 2019

DOI :

10.3791/59915-v

July 26th, 2019

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Jenny Lazarus¹, Yagiz Akiska¹, Mirna Perusina Lanfranca¹, Lawrence Delrosario¹, Lei Sun¹, Daniel Long², Jiaqi Shi³, Howard Crawford², Marina P. Di Magliano¹, Weiping Zou¹, Timothy Frankel¹

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

Podsumowanie

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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Multiplex Fluorescent Immunohistochemistry

Cell Identification

Spatial Analysis

Tumor Microenvironment

Staining Process

Antigen Retrieval

Primary Antibody Diluent

DAPI Working Solution

Deparaffinizing

Rehydrating Slides

Rozdziały w tym wideo

0:04

Title

0:55

Deparaffinization and Rehydration

2:24

Slide Staining

5:05

DAPI Application

5:55

Imaging of Monoplexes and Multiplex Preparation

7:18

Results: Monoplex and Multiplex Optimization

8:39

Conclusion

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