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A Rapid Method for Multispectral Fluorescence Imaging of Frozen Tissue Sections
- Source :
- J Vis Exp
- Publication Year :
- 2020
- Publisher :
- MyJove Corporation, 2020.
-
Abstract
- Multispectral fluorescence imaging on formalin-fixed paraffin-embedded (FFPE) tissues enables the detection of multiple markers in a single tissue sample that can provide information about antigen coexpression and spatial distribution of the markers. However, a lack of suitable antibodies for formalin-fixed tissues may restrict the nature of markers that can be detected. In addition, the staining method is time-consuming. Here we describe a rapid method to perform multispectral fluorescence imaging on frozen tissues. The method includes the fluorophore combinations used, detailed steps for the staining of mouse and human frozen tissues, and the scanning, acquisition, and analysis procedures. For staining analysis, a commercially available semiautomated multispectral fluorescence imaging system is used. Through this method, up to six different markers were stained and detected in a single frozen tissue section. The machine learning analysis software can phenotype cells that can be used for quantitative analysis. The method described here for frozen tissues is useful for the detection of markers that cannot be detected in FFPE tissues or for which antibodies are not available for FFPE tissues.
- Subjects :
- Fluorescence-lifetime imaging microscopy
Tissue Fixation
Fluorophore
General Immunology and Microbiology
Chemistry
General Chemical Engineering
General Neuroscience
Multispectral image
Tissue sample
Fluorescence
Article
General Biochemistry, Genetics and Molecular Biology
Staining
Mice
chemistry.chemical_compound
Multiple markers
Animals
Frozen Sections
Humans
Analysis software
Frozen tissue
Biomedical engineering
Subjects
Details
- ISSN :
- 1940087X
- Database :
- OpenAIRE
- Journal :
- Journal of Visualized Experiments
- Accession number :
- edsair.doi.dedup.....725860ec20b447305bed6ee563ceb922