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Biomarkers are important tools in medicine, which can be used for monitoring disease progression and response to therapy. One of the main problems in rare lysosomal storage diseases is that there are over 70 different diseases, all with different biochemical storage profiles. Developing biochemical biomarkers therefore requires an individual assay per disease/subgroup of diseases. An alternative approach is to develop an assay that is independent of the specific macromolecules stored. This chapter discusses an assay that may serve as a universal biomarker for these diseases and measures the expansion of the late endosomal/lysosomal system. We have developed an assay that takes advantage of a commercially available late endosomal/lysosomal probe, LysoTracker, which becomes trapped in the acidic compartment of cells and emits a fluorescent signal that can be detected using flow cytometry. In this chapter, we detail the methodology behind this assay and discuss the factors that need to be considered when establishing this assay in clinical and research settings.

Original publication

DOI

10.1016/bs.mcb.2014.10.027

Type

Journal article

Journal

Methods Cell Biol

Publication Date

2015

Volume

126

Pages

331 - 347

Keywords

Biomarker, Cellular assay, Flow cytometry, LysoTracker, Lysosomal storage disease, Lysosome, Relative lysosomal volume, Animals, Antigens, CD19, B-Lymphocytes, Humans, Lysosomal Storage Diseases, Lysosomes, Organelle Size