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Journal Article

A new probe for super-resolution imaging of membranes elucidates trafficking pathways.

MPS-Authors
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Kamin,  D.
Department of NanoBiophotonics, MPI for Biophysical Chemistry, Max Planck Society;

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2116939.pdf
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Supplementary Material (public)

2116939_Suppl_1.pdf
(Supplementary material), 2MB

2116939_Suppl_2.zip
(Supplementary material), 7KB

2116939_Suppl_3.zip
(Supplementary material), 5KB

2116939_Suppl_4.zip
(Supplementary material), 13KB

Citation

Revelo, N. H., Kamin, D., Truckenbrodt, S., Wong, A. B., Reuter-Jessen, K., Reisinger, E., et al. (2014). A new probe for super-resolution imaging of membranes elucidates trafficking pathways. Journal of Cell Biology, 205(4), 591-606. doi:10.1083/jcb.201402066.


Cite as: http://hdl.handle.net/11858/00-001M-0000-0025-A918-3
Abstract
The molecular composition of the organelles involved in membrane recycling is difficult to establish as a result of the absence of suitable labeling tools. We introduce in this paper a novel probe, named membrane-binding fluorophore-cysteine-lysine-palmitoyl group (mCLING), which labels the plasma membrane and is taken up during endocytosis. It remains attached to membranes after fixation and permeabilization and can therefore be used in combination with immunostaining and super-resolution microscopy. We applied mCLING to mammalian-cultured cells, yeast, bacteria, primary cultured neurons, Drosophila melanogaster larval neuromuscular junctions, and mammalian tissue. mCLING enabled us to study the molecular composition of different trafficking organelles. We used it to address several questions related to synaptic vesicle recycling in the auditory inner hair cells from the organ of Corti and to investigate molecular differences between synaptic vesicles that recycle actively or spontaneously in cultured neurons. We conclude that mCLING enables the investigation of trafficking membranes in a broad range of preparations.