Multispectral Fluorescence Imaging Allows for Distinctive Topographic Assessment and Subclassification of Tumor-Infiltrating and Surrounding Immune Cells.
IF: 0
Cited by: 11


Histomorphology has significantly changed over the last decades due to technological achievements in immunohistochemistry (IHC) for the visualization of specific proteins and in molecular pathology, particularly in the field of in situ hybridization of small oligonucleotides and amplification of DNA and RNA amplicons. With an increased availability of suitable methods, the demands regarding the observer of histomorphological slides were the supply of complex quantitative data as well as more information about protein expression and cell-cell interactions in tissue sections. Advances in fluorescence-based multiplexed IHC techniques, such as multispectral imaging (MSI), allow the quantification of multiple proteins at the same tissue section. In histopathology, it is a well-known technique for over a decade yet harboring serious problems concerning quantitative preciseness and tissue autofluorescence of multicolor staining when using formalin-fixed, paraffin-embedded (FFPE) tissue specimen. In recent years, milestones in tissue preparation, fluorescent dyes, hardware imaging, and software analysis were achieved including automated tissue segmentation (e.g., tumor vs. stroma) as well as in cellular and subcellular multiparameter analysis.This chapter covers the role that MSI plays in anatomic pathology for the analysis of FFPE tissue sections, discusses the technical aspects of MSI, and provides a review of its application in the characterization of immune cell infiltrates and beyond regarding its prognostic and predictive value and its use for guidance of clinical decisions for immunotherapeutic strategies.


Fluorescence microscopy
Immune cells
Multispectral imaging

MeSH terms

Antineoplastic Agents, Immunological
Biomarkers, Tumor
Fluorescent Antibody Technique
Image Processing, Computer-Assisted
Microscopy, Fluorescence
Neoplasms, Experimental
Paraffin Embedding
Tissue Fixation


Wickenhauser, Claudia
Bethmann, Daniel
Feng, Zipei
Jensen, Shawn M
Ballesteros-Merino, Carmen
Massa, Chiara
Steven, Andre
Bauer, Marcus
Kaatzsch, Peter
Pazaitis, Nikolaos
Toma, Georgiana
Bifulco, Carlo B
Fox, Bernard A
Seliger, Barbara

Recommend literature