Experimental Perspectives on Direct Visualization of Endosomal Rupture

Endosomal escape continues to be a limiting factor in the therapeutic use of nanomaterials. Assays to visualize endosomal escape often do not decouple the endosomal/lysosomal disruption from the release of payload into the cytosol. Here, we discuss three approaches to directly probe endosomal/lysoso...

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Bibliographic Details
Published in:Chembiochem : a European journal of chemical biology 2021-12, Vol.22 (23), p.3277-3282
Main Authors: Day, Rachael A., Sletten, Ellen M.
Format: Article
Language:English
Subjects:
Calcein
Cytosol
Cytosol - chemistry
Dilution
drug delivery
Endosomes
Endosomes - chemistry
Fluoresceins - chemistry
Genetic engineering
Humans
Internalization
lysosome size
Lysosomes
Lysosomes - chemistry
nanoemulsions
Nanomaterials
Nanotechnology
Osmosis
Osmotic Pressure
osmotic rupture
photochemical internalization
Photochemical Processes
Photochemicals
Rupture
Rupturing
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Summary:Endosomal escape continues to be a limiting factor in the therapeutic use of nanomaterials. Assays to visualize endosomal escape often do not decouple the endosomal/lysosomal disruption from the release of payload into the cytosol. Here, we discuss three approaches to directly probe endosomal/lysosomal rupture: calcein dye dilution, lysosome size quantification and endosome/lysosome membrane integrity visualized with a genetically engineered cell line. We apply the three assays to endosomes/lysosomes ruptured via osmotic pressure and photochemical internalization. Endosomal/lysosomal disruption can be directly visualized via lysosome size or genetically engineered cell lines.
ISSN:1439-4227
1439-7633
DOI:10.1002/cbic.202100379