title: Visualizing Lysosomal Membrane Permeabilization by Fluorescent Dextran Release.
abstract: Lysosomal membrane permeabilization (LMP) is an effective programmed cell death pathway triggered in response to a variety of cytotoxic stimuli and cellular conditions. In the method presented here, LMP is monitored by first taking advantage of the steady endocytic capacity of cells to load fluorescent dextran into lysosomes, and then simply observing the translocation of lysosomally localized dextran into the cytosol after an LMP-inducing insult. Fluorescent dextran in healthy cells appears in punctate structures representing intact lysosomes, whereas after LMP, a diffuse staining pattern throughout the cytoplasm is observed. Using this method, LMP can be followed in real time using time-lapse imaging. The size of pores formed in the membrane during LMP by size exclusion can also be determined using dextrans of different sizes and colors.
authors.affiliation: Unit for Cell Death and Metabolism, Center for Autophagy, Recycling and Disease, Danish Cancer Society Research Center, DK-2100, Copenhagen, Denmark.
authors.forename: Anne-Marie
authors.initials: AM
authors.lastname: Ellegaard
authors.affiliation: Unit for Cell Death and Metabolism, Center for Autophagy, Recycling and Disease, Danish Cancer Society Research Center, DK-2100, Copenhagen, Denmark.
authors.forename: Marja
authors.initials: M
authors.lastname: Jäättelä
authors.affiliation: Unit for Cell Death and Metabolism, Center for Autophagy, Recycling and Disease, Danish Cancer Society Research Center, DK-2100, Copenhagen, Denmark.
authors.forename: Jesper
authors.initials: J
authors.lastname: Nylandsted
mesh_terms: D000818:Animals; D002460:Cell Line; D003600:Cytosol / Q000737:chemistry*; D003911:Dextrans / Q000032:analysis*; D005456:Fluorescent Dyes / Q000032:analysis*; D006801:Humans; D007425:Intracellular Membranes / Q000502:physiology*; D008247:Lysosomes / Q000737:chemistry*; D010539:Permeability; D059008:Time-Lapse Imaging
chemical_list: D003911:Dextrans; D005456:Fluorescent Dyes