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This volume provides a wide range of methods to discover and assess lysosome structure and function are provided. Chapters detail the essential methodologies and giving expert insight to key features with notes pertaining to key elements of the methods. Written in the highly successful Methods in Molecular Biology series format, the chapters include brief introductions to the material, lists of necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and a Notes section which highlights tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Lysosomes: Methods and Protocols aims to be comprehensive guide for researchers in the field.
Determining lysosomal enzyme activity using flurogenic probes.- Lysosomal pH measurements via dual-imaging microscopy.- Assessing perturbations in lysosome health by flow cytometry in iPSC-derived human neuron cultures.- Label-Free Imaging of Lysosomes in Living Cells by Photothermal Microscopy.- Monitoring autophagosome biogenesis and degradation in mammalian cells using LC3B blotting.- Measurement of physiological autophagic flux in the human peripheral blood mononuclear cell pool.- Lysosome enrichment using superparamagnetic iron oxide nanoparticles (SPIONs).- Lysosomal immunoprecipitation (LysoIP) for metabolomic and proteomic analyses.- Mass spectrometry-based proteomic analysis of lysosome-enriched fractions.- Analyzing Endolysosome-Mitochondria Membrane Contact Sites by Thin Section TEM of Human Fibroblasts.- Assessment of lysosome trafficking in iPSC-derived neurons utilizing time-lapse videos and TrackMate ImageJ plugin software.- Generation of donor-specific iPSC for modelling lysosomal storage disorders.- Using CMAC Staining for Vacuole Characterization in Yeast.- Modelling lysosomal disease in Dictyostelium discoideum: Examining the trafficking and secretion of lysosomal enzymes.- Bioenergetic profiling applied to a zebrafish model of Neuronal Ceroid Lipofuscinosis, a lysosomal storage disorder.- Immunostaining methods for lysosomal storage disorder research.
