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Laser Capture and Single Cell Genotyping from Frozen Tissue Sections
Thomas Kroneis1,2, Jody Ye3, Kathleen Gillespie3
1Institute of Cell Biology, Histology & Embryology, Medical University of Graz, Harrachgasse 21/7, 8010, Graz, Austria. thomas.kroneis@medunigraz.at.
Methods in Molecular Biology (Clifton, N.J.)
|December 15, 2015
Summary
This study presents a method for isolating single cells from tissue sections using laser microdissection and pressure catapulting (LMPC). This technique enables precise genetic analysis, crucial for applications like tumor cell identification and chimerism studies in pancreas tissue.
Area of Science:
- Genomics
- Molecular Biology
- Cellular Biology
Background:
- Increasing need for single-cell genetic analysis from tissue sections.
- Applications include tumor cell analysis and studying pancreatic cells in type 1 diabetes or chimerism.
Purpose of the Study:
- To describe a protocol for isolating cells from tissue sections for genetic analysis.
- To enable discrimination of haplo-identical cells (fetal and maternal) in pancreas tissue.
Main Methods:
- Cryo-sectioning of snap-frozen tissues mounted on membrane-coated slides.
- Laser microdissection and pressure catapulting (LMPC) for cell isolation.
- 17-plex PCR amplification of short tandem repeats (STR) and amelogenin locus for DNA profiling.
Main Results:
- Successful isolation of individual cells from tissue sections.
- Demonstrated capability for DNA profiling using STR and amelogenin loci.
- Potential for discriminating between haplo-identical cells.
Conclusions:
- The described protocol provides a robust method for single-cell isolation and genetic analysis from tissue sections.
- This technique is valuable for various research areas, including oncology and transplantation immunology.
- Further DNA profiling methods can be integrated for enhanced analysis.
