Protocol for sorting cells from mouse brains labeled with mosaic analysis with double markers by flow cytometry
Nicole Amberg1, Giselle Cheung1, Simon Hippenmeyer1
1Institute of Science and Technology Austria (ISTA), Am Campus 1, 3400 Klosterneuburg, Austria.
STAR Protocols
|December 9, 2023
Summary
This study details a protocol for isolating mosaic analysis with double markers (MADM)-labeled cells from mice using fluorescence-activated cell sorting (FACS). This method yields high-quality cells for molecular analysis across various developmental stages.
Area of Science:
- Developmental Biology
- Genetics
- Cell Biology
Background:
- Mosaic analysis with double markers (MADM) technology allows for genetic mosaic generation in mice.
- High-resolution phenotyping at the single-cell level is crucial for understanding complex biological processes.
Purpose of the Study:
- To present a high-yield protocol for isolating MADM-labeled cells.
- To enable downstream molecular analyses of genetically mosaic tissues in mice.
Main Methods:
- Generation of MADM-labeled mice.
- Perfusion, single-cell suspension, and debris removal procedures.
- Fluorescence-activated cell sorting (FACS) for cell isolation and downstream analysis.
Main Results:
- A detailed protocol for isolating MADM-labeled cells with high yield.
- The protocol is applicable to embryonic to adult mouse tissues.
- Facilitates robust downstream molecular analyses.
Conclusions:
- The presented FACS-based protocol effectively isolates MADM-labeled cells.
- This method supports detailed molecular investigation of genetic mosaics in mice.
- The protocol is adaptable for various developmental stages.
