The acidic environment of the lysosomal compartment provides an optimal setting for acid hydrolases and is essential for fusion/fission and cargo sorting of the endolysosomal compartment. Maintenance of lysosomal acidity is essential for disease avoidance. CD BioSciences' expertise in lysosomes provides strong support for the assessment of lysosomal pH service. Our advanced technology platform will bring you an efficient and cost-effective service that will ensure your 100% satisfaction.
Introduction of Lysosomal pH
Lysosomes contain hydrolases that promote the breakdown of proteins, lipids and polysaccharides. Lysosomes maintain cellular homeostasis by generating a highly acidic environment at pH 4.5-5.0 and by accommodating hydrolases that degrade phagocytosed biomolecules. It has been shown that lysosomal acidification is achieved through the activity of vesicular ATPases (V-ATPases). Dysregulated lysosomal acidification and defective lysosomal-organelle fusion lead to impaired endocytosis, autophagic degradation, macromolecular biogenesis and translocation and are observed in protein-based neurodegenerative diseases, metabolic disorders and immune disorders. Lysosomal acidification dysfunction also affects the activity of other organelles, such as mitochondria, leading to increased production of reactive oxygen species and inflammatory cytokines, which contribute to the pathogenesis of inflammatory diseases, cancer, and infectious diseases. Thus, lysosomal pH is an attractive and targetable site for therapeutic intervention in disease.
Fig. 1. Determinants of lysosomal pH. (DiCiccio J E, et al., 2011)
Our Lysosomal pH Assessment Services
There is a wealth of recent data output on lysosomal acidification. Fluorescent pH probes have proven to be an effective tool for studying the role of intracellular pH in a variety of physiological and pathological processes. CD BioSciences has developed a protocol for analyzing lysosomal pH in cultured cells using fluorescent probes as well as a dual emission ratio technique suitable for flow cytometry. Here, our engineers build multiple strategies to determine lysosomal pH.
- Acidic Dyes Are Used to Estimate Lysosomal pH
This is a simple method that provides indirect and qualitative measurements of pH. - Qualitative Assessment of Lysosomal Acidification Using LysoTracker
The LysoTracker probe is suitable for labeling lysosomes as well as late endosomes, and the fluorescent signal is not used as a measure of lysosomal pH. - Ratiometric Assessment of Lysosomal pH Using Exogenous Probes
We use the LysoSensor probe to label lysosomes. This approach allows for ratiometric measurement of lysosomal pH using dual-wavelength fluorescence-based analysis, and also allows for ratiometric assessment of lysosomal pH using pulse-tracking liquid phase labeling. - Ratiometric Assessment of Lysosomal pH Using Genetically Encoded Probes
We use a genetically encoded pH lysosomal biosensor for fluorescence microscopy imaging. This approach allows for robust measurements of lysosomal pH over extended periods of time ranging from hours to days.
Features of Our Services
- Lysosomal targeting probes are usually weak bases that are selectively focused in acidic organelles due to protonation and can ensure retention in the lysosome during pH changes.
- The ratio of emission intensities detected at wavelengths will vary with pH, but will not be affected by the number of probes available.
- Flow cytometry can be performed on a large number of cells that are quickly measured and analyzed.
- The pH of the lysosome is determined by establishing a standard curve of FITC-dextrose obtained from the cells.
- Peripheral and proximal nuclear lysosomal subpopulations can be visualized using genetically encoded ratio probes.
- The fluorescence ratios of pH-sensitive and pH-insensitive fluorophores can be measured by flow cytometry.
Our lysosomal pH assessment service provides you with the most appropriate and rigorous method to measure lysosomal pH. Our highly skilled and dedicated scientific staff ensures that the most appropriate method and technology is selected for each specialized lysosomal project. If you have any special requirements about our services, please feel free to contact us. We are looking forward to working together with your attractive projects.
References
- DiCiccio J E, Steinberg B E. (2011) Lysosomal pH and analysis of the counter ion pathways that support acidification[J]. Journal of General Physiology. 137(4): 385-390.
- Eriksson I, Öllinger K, Appelqvist H. (2017) Analysis of Lysosomal pH by Flow Cytometry Using FITC-Dextran Loaded Cells. Methods Mol Biol. 1594:179-189.