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Putting the More back in Morphology: Quantitating Multiple Protein Expressions in Intact Tissue Sections
Author(s) -
Mansfield James Robert,
Hoyt Clifford
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.657.3
Subject(s) - autofluorescence , multispectral image , segmentation , computer science , fluorescence lifetime imaging microscopy , flow cytometry , biomedical engineering , computational biology , fluorescence microscope , sensitivity (control systems) , microscopy , pathology , pattern recognition (psychology) , artificial intelligence , fluorescence , biology , medicine , microbiology and biotechnology , physics , optics , electronic engineering , engineering
Biological systems are complex; multiparameter detection methods such as expression arrays and flow cytometry make this apparent. In tissues it is important not just to measure the average expression of molecules, but also their spatial distribution, while preserving architectural features. Such high‐resolution molecular imaging is challenging, especially when signals are co‐localized. In fluorescence, sensitivity and quantitation can be compromised by autofluorescence, which is commonly present in formalin‐fixed tissues. Imaging challenges can be addressed using multispectral imaging. On a microscope, at least 6 markers can be simultaneously detected and imaged, and sensitivity can be increased by two orders of magnitude, depending on relative levels of specific label and autofluorescence. For analysis, we have developed a train‐by‐example software platform that can be trained to separate image regions into appropriate classes (“cancer”, “inflammation”, e.g.) and to accurately quantitate their areas. When combined with multispectral imaging and cellular segmentation tools to extract data automatically from cellular and sub‐cellular compartments it can provide the information necessary for designing and testing targeted diagnostic and therapeutic reagents.