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Protocol for labeling and fixation of intact lysosomes with esterified amino acid analogs to assess lysosomal expansion in living eukaryotic cells

The lysosomal compartment is a key hub for cell metabolism, and morphological alterations have been described in several pathological conditions. Here, we describe the use of amino acid analogs modified by the presence of a methyl ester group that accumulates within lysosomes. This generates an intr...

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Autores principales: Scerra, Gianluca, Caporaso, Maria Gabriella, Renna, Maurizio, D’Agostino, Massimo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8564676/
https://www.ncbi.nlm.nih.gov/pubmed/34755119
http://dx.doi.org/10.1016/j.xpro.2021.100916
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author Scerra, Gianluca
Caporaso, Maria Gabriella
Renna, Maurizio
D’Agostino, Massimo
author_facet Scerra, Gianluca
Caporaso, Maria Gabriella
Renna, Maurizio
D’Agostino, Massimo
author_sort Scerra, Gianluca
collection PubMed
description The lysosomal compartment is a key hub for cell metabolism, and morphological alterations have been described in several pathological conditions. Here, we describe the use of amino acid analogs modified by the presence of a methyl ester group that accumulates within lysosomes. This generates an intraluminal osmotic effect able to trigger a rapid and selective expansion of the lysosomal compartment within 2 h of treatment. We also present protocols to preserve lysosomal morphology, which yields a more accurate size measurement. For complete details on the use and execution of this protocol, please refer to Scerra et al. (2021).
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spelling pubmed-85646762021-11-08 Protocol for labeling and fixation of intact lysosomes with esterified amino acid analogs to assess lysosomal expansion in living eukaryotic cells Scerra, Gianluca Caporaso, Maria Gabriella Renna, Maurizio D’Agostino, Massimo STAR Protoc Protocol The lysosomal compartment is a key hub for cell metabolism, and morphological alterations have been described in several pathological conditions. Here, we describe the use of amino acid analogs modified by the presence of a methyl ester group that accumulates within lysosomes. This generates an intraluminal osmotic effect able to trigger a rapid and selective expansion of the lysosomal compartment within 2 h of treatment. We also present protocols to preserve lysosomal morphology, which yields a more accurate size measurement. For complete details on the use and execution of this protocol, please refer to Scerra et al. (2021). Elsevier 2021-10-27 /pmc/articles/PMC8564676/ /pubmed/34755119 http://dx.doi.org/10.1016/j.xpro.2021.100916 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Scerra, Gianluca
Caporaso, Maria Gabriella
Renna, Maurizio
D’Agostino, Massimo
Protocol for labeling and fixation of intact lysosomes with esterified amino acid analogs to assess lysosomal expansion in living eukaryotic cells
title Protocol for labeling and fixation of intact lysosomes with esterified amino acid analogs to assess lysosomal expansion in living eukaryotic cells
title_full Protocol for labeling and fixation of intact lysosomes with esterified amino acid analogs to assess lysosomal expansion in living eukaryotic cells
title_fullStr Protocol for labeling and fixation of intact lysosomes with esterified amino acid analogs to assess lysosomal expansion in living eukaryotic cells
title_full_unstemmed Protocol for labeling and fixation of intact lysosomes with esterified amino acid analogs to assess lysosomal expansion in living eukaryotic cells
title_short Protocol for labeling and fixation of intact lysosomes with esterified amino acid analogs to assess lysosomal expansion in living eukaryotic cells
title_sort protocol for labeling and fixation of intact lysosomes with esterified amino acid analogs to assess lysosomal expansion in living eukaryotic cells
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8564676/
https://www.ncbi.nlm.nih.gov/pubmed/34755119
http://dx.doi.org/10.1016/j.xpro.2021.100916
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