In Vivo Analysis of Dendritic Cell Clonality
1Immunobiology Laboratory, The Francis Crick Institute, London, UK. mdmc2@cam.ac.uk.
Insights
This study presents a multicolor fate mapping technique to track conventional dendritic cell (cDC) progeny in tissues. The method aids in analyzing cell distribution and understanding cDC clonality using advanced microscopy.
Area of Science:
- Immunology
- Cell Biology
- Microscopy
Background:
- Confocal fluorescence microscopy is vital for analyzing tissue architecture and cell distribution.
- Multicolor fate mapping combined with microscopy enables the study of cell lineage and clonal relationships.
Purpose of the Study:
- To describe a multicolor fate mapping mouse model and microscopy technique.
- To trace the progeny of conventional dendritic cell (cDC) progenitors in various tissues.
- To analyze cDC clonality and understand their behavior within tissues.
Main Methods:
- Development of a multicolor fate mapping mouse model.
- Application of confocal fluorescence microscopy for high-resolution imaging.
- Focus on imaging methodologies for tracing cell lineages.
Main Results:
- The described technique allows for the visualization and tracing of conventional dendritic cell (cDC) progeny.
- The method facilitates the analysis of cell clusters and informs on clonal relationships.
- Imaging protocols are detailed for studying cDC clonality.
Conclusions:
- This chapter provides a robust method for studying conventional dendritic cell (cDC) development and behavior.
- The described imaging techniques are crucial for understanding tissue-specific cDC clonality.
- The methodology supports further research into immune cell dynamics and tissue organization.
Abstract:
Confocal fluorescence microscopy is commonly used for the analysis of tissue architecture and cell distribution (Paddock, Confocal microscopy: methods and protocols. Methods in molecular biology. Humana Press, New York, pp 1-388, 2013). When combined with multicolor fate mapping of cell precursors, it allows for analysis of single-color cell clusters, which in turn informs on the clonal relationship of cells in tissues (Snippert et al, Cell 143:134-144. https://doi.org/10.1016/j.cell.2010.09.016 , 2010). In this chapter, I describe a multicolor fate mapping mouse model and microscopy technique to trace the progeny of conventional dendritic cell (cDC, (Cabeza-Cabrerizo et al, Annu Rev Immunol 39:131. https://doi.org/10.1146/annurev-immunol-061020-053707 , 2021)) progenitors in different tissues and analyze cDC clonality. The chapter is focused on imaging methods rather than image analysis, although the software used to quantify cluster formation is also introduced.
