Multiplex IHC Analysis of FFPE Slides

Five FFPE slides from specimens, with observed TLS in the H&E-stained sections, were subjected to multispectral immunohistochemical (mIHC) staining29 30 (link) using the Opal color kit (PerkinElmer, Hopkinton, Massachusetts, USA), according to the manufacturer’s instructions, to provide simultaneous detection of adaptive immune system cell types, such as CD4⁺ T cells, CD8⁺ T cells, CD20⁺ B cells, and CD45RO⁺ memory T cells. Cell nuclei were stained with DAPI. Multiplexed color slides were imaged with a PerkinElmer Vectra automated multispectral microscope at 100× magnification and analyzed using PerkinElmer inForm Analysis Software.

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Zhang W.H., Wang W.Q., Han X., Gao H.L., Xu S.S., Li S., Li T.J., Xu H.X., Li H., Ye L.Y., Lin X., Wu C.T., Long J., Yu X.J, & Liu L. (2020). Infiltrating pattern and prognostic value of tertiary lymphoid structures in resected non-functional pancreatic neuroendocrine tumors. Journal for Immunotherapy of Cancer, 8(2), e001188.

Publication 2020

inForm Analysis Software

Adaptive B cells Cd4 t cells Cd45ro Cd8 t cells Cell nuclei Cell types Dapi Immune system Memory t cells Microscope Opal Vectra

Corresponding Organization : Shanghai Cancer Institute

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Variable analysis

independent variables

MIHC staining using the Opal color kit

dependent variables

Simultaneous detection of adaptive immune system cell types (CD4+ T cells, CD8+ T cells, CD20+ B cells, CD45RO+ memory T cells)

control variables

Five FFPE slides from specimens with observed TLS in the H&E-stained sections
Cell nuclei stained with DAPI

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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