Research ResourcesIMMUNOTHERAPY

In-depth tissue profiling using multiplexed immunohistochemical consecutive staining on single slide

See allHide authors and affiliations

Science Immunology  14 Jul 2016:
Vol. 1, Issue 1, pp. aaf6925
DOI: 10.1126/sciimmunol.aaf6925

You are currently viewing the abstract.

View Full Text

Log in to view the full text

Log in through your institution

Log in through your institution

Putting immune cells on the map

Multiplexed flow cytometry allows for careful characterization of the complexity of immune cells. However, these cells do not act in isolation, and multiplexed analyses do not extend to spatial visualization of cells in tissue microenvironments. Remark et al. have combined immunohistochemistry with virtual landscaping to generate maps of immune cells in tumors stained with up to 10 distinct antibodies. Rather than using filters to resolve chromogens, they use multiple rounds of staining and destaining, followed by virtual color assignment. This simple yet powerful technique can be used to chart maps of cells in any tissue.


Despite remarkable recent achievements of immunotherapy strategies in cancer treatment, clinical responses remain limited to subsets of patients. Predictive markers of disease course and response to immunotherapy are urgently needed. Recent results have revealed the potential predictive value of immune cell phenotype and spatial distribution at the tumor site, prompting the need for multidimensional immunohistochemical analyses of tumor tissues. To address this need, we developed a sample-sparing, highly multiplexed immunohistochemistry technique based on iterative cycles of tagging, image scanning, and destaining of chromogenic substrate on a single slide. This assay, in combination with a newly developed automated digital landscaping solution, democratizes access to high-dimensional immunohistochemical analyses by capturing the complexity of the immunome using routine pathology standards. Applications of the method extend beyond cancer to screen and validate comprehensive panels of tissue-based prognostic and predictive markers, perform in-depth in situ monitoring of therapies, and identify targets of disease.

View Full Text

Stay Connected to Science Immunology