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Purification of Specific Cell Population by Fluorescence Activated Cell Sorting FACS
Published on: July 10, 2010
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Purification of Synaptosome Populations Using Fluorescence-Activated Synaptosome Sorting
Elisa Luquet1,2, Christoph Biesemann3, Annie Munier4
1Interdisciplinary Institute for Neuroscience, University Bordeaux, CNRS, UMR 5297, F-33000, Bordeaux, France.
Methods in Molecular Biology (Clifton, N.J.)
|December 13, 2016
Summary
Researchers developed Fluorescence Activated Synaptosome Sorting to purify brain cell components. This method significantly enriches specific synaptosomes, improving molecular analysis of the central nervous system.
Area of Science:
- Neurobiology
- Cellular Neuroscience
- Biochemistry
Background:
- Subcellular fractionation is crucial for analyzing central nervous system molecular structure.
- Traditional methods like synaptosome preparation lack specificity and purity.
- Brain tissue complexity necessitates refined isolation techniques.
Purpose of the Study:
- To present a detailed protocol for Fluorescence Activated Synaptosome Sorting (FASS).
- To improve the specificity and purity of isolated synaptosomes for research.
Main Methods:
- Extension of conventional synaptosome preparation.
- Utilizing cell sorting for purification of fluorescent synaptosomes.
- Employing VGLUT1venus reporter for excitatory synaptosome identification.
Main Results:
- Achieved steep enrichment of fluorescent excitatory VGLUT1venus synaptosomes.
- Successfully isolated synaptosomes containing presynaptic and postsynaptic elements.
- Demonstrated strong depletion of neuronal and glial contaminants.
Conclusions:
- Fluorescence Activated Synaptosome Sorting offers enhanced purity and specificity.
- This method provides a more refined tool for studying central nervous system components.
- Improved synaptosome isolation facilitates advanced molecular and functional analyses.
