Methods to Detect Loss of Lysosomal Membrane Integrity
Sonja Aits1,2
1Cell Death and Lysosomes Group, Experimental Neuroinflammation Laboratory, Department of Experimental Medical Science, Lund University, Lund, Sweden. sonja.aits@med.lu.se.
Methods in Molecular Biology (Clifton, N.J.)
|January 6, 2019
Summary
Lysosomal membrane permeabilization (LMP) is detected using the galectin puncta assay. This sensitive method visualizes galectin translocation to damaged lysosomes in live or fixed cells.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Lysosomal membrane permeabilization (LMP) is a critical event in various cell death pathways.
- Existing methods for LMP detection have limitations in scope or ease of use.
Purpose of the Study:
- To provide detailed protocols for the galectin puncta formation assay for detecting LMP.
- To introduce an alternative assay based on fluorescent dextran release for parallel assessment of LMP.
Main Methods:
- Detailed protocols for galectin puncta formation assay in fixed and live cells.
- Methodology for fluorescent dextran release assay from damaged lysosomes.
Main Results:
- The galectin puncta formation assay is highly sensitive and applicable to both live and fixed cells.
- The assay allows visualization of galectin translocation to damaged lysosomes, indicated by a shift from uniform to punctate staining.
Conclusions:
- The galectin puncta formation assay is presented as a robust and versatile method for detecting LMP.
- The provided protocols facilitate standardized and reliable assessment of lysosomal membrane integrity in cellular studies.
Keywords:
Cell deathDextranEGFP-galectin-3GalectinImmunocytochemistryLysosomal membrane permeabilizationLysosomeLysosome-associated membrane proteinMicroscopyMore Related Videos
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