Improved multiplex immunohistochemistry for immune microenvironment evaluation of mouse formalin-fixed, paraffin-embedded tissues

Noah Sorrelle, Debolina Ganguly, Adrian T.A. Dominguez, Yuqing Zhang, Huocong Huang, Lekh N. Dahal, Natalie Burton, Arturas Ziemys, Rolf A Brekken

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Immune profiling of tissue through multiplex immunohistochemistry is important for the investigation of immune cell dynamics, and it can contribute to disease prognosis and evaluation of treatment response in cancer patients. However, protocols for mouse formalinfixed, paraffin-embedded tissue have been less successful. Given that formalin fixation and paraffin embedding remains the most common preparation method for processing mouse tissue, this has limited the options to study the immune system and the impact of novel therapeutics in preclinical models. In an attempt to address this, we developed an improved immunohistochemistry protocol with a more effectiveAg-retrieval buffer.We also validated 22 Abs specific formouse immune cellmarkers to distinguish B cells,T cells, NKcells, macrophages, dendritic cells, and neutrophils. In addition, we designed and tested novel strategies to identify immune cells for which unique Abs are currently not available. Last, in the 4T1 model of breast cancer, we demonstrate the utility of our protocol andAb panels in the quantitation and spatial distribution of immune cells.

Original languageEnglish (US)
Pages (from-to)292-299
Number of pages8
JournalJournal of Immunology
Volume202
Issue number1
DOIs
StatePublished - Jan 1 2019

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Paraffin
Formaldehyde
Immunohistochemistry
Paraffin Embedding
Dendritic Cells
Immune System
Buffers
Neutrophils
B-Lymphocytes
Macrophages
Breast Neoplasms
T-Lymphocytes
Therapeutics
Neoplasms

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Improved multiplex immunohistochemistry for immune microenvironment evaluation of mouse formalin-fixed, paraffin-embedded tissues. / Sorrelle, Noah; Ganguly, Debolina; Dominguez, Adrian T.A.; Zhang, Yuqing; Huang, Huocong; Dahal, Lekh N.; Burton, Natalie; Ziemys, Arturas; Brekken, Rolf A.

In: Journal of Immunology, Vol. 202, No. 1, 01.01.2019, p. 292-299.

Research output: Contribution to journalArticle

Sorrelle, Noah ; Ganguly, Debolina ; Dominguez, Adrian T.A. ; Zhang, Yuqing ; Huang, Huocong ; Dahal, Lekh N. ; Burton, Natalie ; Ziemys, Arturas ; Brekken, Rolf A. / Improved multiplex immunohistochemistry for immune microenvironment evaluation of mouse formalin-fixed, paraffin-embedded tissues. In: Journal of Immunology. 2019 ; Vol. 202, No. 1. pp. 292-299.
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