Background of Lysosomal Galectin Puncta
Studies on the involvement of lysosomes in cell death are limited due to the poor sensitivity or the susceptibility to artifacts of existing methods for detecting lysosomal membrane permeability (LMP). Induction of LMP triggers the rapid translocation of Galectin 3/LGALS3 from the cytosol to the lysosomal lumen, making it an important marker of LMP. In addition, Galectin 3 can also be recruited to damaged intima/phagosomal membranes. Galectin is a soluble carbohydrate-binding lectin. Studies have shown that cytoplasmic LGALS3/galactosin-3 plays an important role in autophagy-mediated defense against Salmonella typhimurium invasion of the host cytoplasm. Therefore, the lysosomal galectin puncta assay opens up new possibilities to study the role of LMPs in cell death and their roles in other cellular processes.
Fig. 1. Galectin puncta assay is suitable for automated analysis of LMP. (Aits S, et al., 2015)
CLysosomal Galectin Puncta Analysis Services
Galectin staining can mark individual leaky lysosomes early in lysosomal cell death and assist in determining the role of LMP in cell death. Over the years, CD BioSciences offers multiple methods to quantify and visualize lysosomal permeability during cell death. Here, we performed the detection of lysosomal membrane permeability by the lysosomal galectin puncta assay, based on the transport of galectin to damaged lysosomes, resulting in a change from uniform to speckled staining patterns.
We typically choose LGALS1 and LGALS3, which are widely expressed and rapidly translocated, and which also provide corresponding high-affinity antibodies. Our engineers have developed a detailed protocol for the detection of galectin puncta in leaky lysosomes to assess LMP, allowing labelling of individual leaky lysosomes in cells and paraffin-embedded tissues.
In addition, we used ectopic fluorescent galectin (LGALS1, LGALS3, LGALS8, or LGALS9) for automated screening and visualization of LMP in live cells and animals.This method is widely welcomed by customers around the world due to its ease of operation and high sensitivity.
Advantages of This Approach
- Galectin puncta assays are faster and more sensitive than methods that monitor cathepsin release.
- Allows detection of LMPs without cell death.
- Allows a subset of damaged lysosomes to be distinguished from other intact lysosomes within the same cell.
- High-throughput imaging platform suitable for combination with other cell viability markers.
- Allows high-throughput detection and quantification of damaged lysosomes by fluorescence microscopy.
CD BioSciences can meet any reasonable needs of our clients, taking time and budget into consideration for you. Our aim is to be customer-centric and to provide the highest quality services to customers. Our customer service representatives are enthusiastic and trustworthy 24 hours a day, 7 days a week. If you are interested in our services, please feel free to contact us for more information or a detailed discussion.
Reference
- Aits S, Kricker J, et al. (2015) Sensitive detection of lysosomal membrane permeabilization by lysosomal galectin puncta assay. Autophagy. 11(8):1408-24.