Multiplexed quantification of autophagic flux by imaging flow cytometry

Léa Montégut; Hui Chen; Gerasimos Anagnostopoulos; Sabrina Spaggiari; Oliver Kepp; Maria Chiara Maiuri; Guido Kroemer; Isabelle Martins

doi:10.1016/bs.mcb.2021.02.005

Multiplexed quantification of autophagic flux by imaging flow cytometry

Léa Montégut, Hui Chen, Gerasimos Anagnostopoulos, Sabrina Spaggiari, Oliver Kepp, Maria Chiara Maiuri, Guido Kroemer, Isabelle Martins

Résultats de recherche: Le chapitre dans un livre, un rapport, une anthologie ou une collection › !!Chapter › Revue par des pairs

2 Citations (Scopus)

Résumé

Imaging flow cytometry allows for the quantitative assessment of fluorescent signals at the subcellular level. Here, we describe the use of a biosensor cell line, namely, U2OS osteosarcoma cells equipped with a fusion protein comprising monomeric red fluorescent protein (mRFP), green fluorescent protein (GFP) and microtubule-associated proteins 1A/1B light chain 3B (best known as LC3), for the assessment of autophagic flux by imaging flow cytometry. We detail all analysis tools required to distinguish autophagosomes (that emit both a red and a green fluorescence) and autolysosomes (that emit a red fluorescence, yet lose the green fluorescent signal) and to quantitate autophagic flux in a convenient fashion.

langue originale	Anglais
titre	Monitoring Vesicular Trafficking in Cellular Responses to Stress - Part B
rédacteurs en chef	Oliver Kepp, Lorenzo Galluzzi
Editeur	Academic Press Inc.
Pages	59-71
Nombre de pages	13
ISBN (imprimé)	9780128244876
Les DOIs	https://doi.org/10.1016/bs.mcb.2021.02.005
état	Publié - 1 janv. 2021

Série de publications

Nom	Methods in Cell Biology
Volume	165
ISSN (imprimé)	0091-679X

Accès au document

10.1016/bs.mcb.2021.02.005

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Contient cette citation

Montégut, L., Chen, H., Anagnostopoulos, G., Spaggiari, S., Kepp, O., Maiuri, M. C., Kroemer, G., & Martins, I. (2021). Multiplexed quantification of autophagic flux by imaging flow cytometry. Dans O. Kepp, & L. Galluzzi (eds.), Monitoring Vesicular Trafficking in Cellular Responses to Stress - Part B (p. 59-71). (Methods in Cell Biology; Vol 165). Academic Press Inc.. https://doi.org/10.1016/bs.mcb.2021.02.005

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abstract = "Imaging flow cytometry allows for the quantitative assessment of fluorescent signals at the subcellular level. Here, we describe the use of a biosensor cell line, namely, U2OS osteosarcoma cells equipped with a fusion protein comprising monomeric red fluorescent protein (mRFP), green fluorescent protein (GFP) and microtubule-associated proteins 1A/1B light chain 3B (best known as LC3), for the assessment of autophagic flux by imaging flow cytometry. We detail all analysis tools required to distinguish autophagosomes (that emit both a red and a green fluorescence) and autolysosomes (that emit a red fluorescence, yet lose the green fluorescent signal) and to quantitate autophagic flux in a convenient fashion.",

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Montégut, L, Chen, H, Anagnostopoulos, G, Spaggiari, S, Kepp, O, Maiuri, MC, Kroemer, G & Martins, I 2021, Multiplexed quantification of autophagic flux by imaging flow cytometry. Dans O Kepp & L Galluzzi (eds), Monitoring Vesicular Trafficking in Cellular Responses to Stress - Part B. Methods in Cell Biology, VOL. 165, Academic Press Inc., p. 59-71. https://doi.org/10.1016/bs.mcb.2021.02.005

Multiplexed quantification of autophagic flux by imaging flow cytometry. / Montégut, Léa; Chen, Hui; Anagnostopoulos, Gerasimos et al.
Monitoring Vesicular Trafficking in Cellular Responses to Stress - Part B. Ed. / Oliver Kepp; Lorenzo Galluzzi. Academic Press Inc., 2021. p. 59-71 (Methods in Cell Biology; Vol 165).

Résultats de recherche: Le chapitre dans un livre, un rapport, une anthologie ou une collection › !!Chapter › Revue par des pairs

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T1 - Multiplexed quantification of autophagic flux by imaging flow cytometry

AU - Montégut, Léa

AU - Chen, Hui

AU - Anagnostopoulos, Gerasimos

AU - Spaggiari, Sabrina

AU - Kepp, Oliver

AU - Maiuri, Maria Chiara

AU - Kroemer, Guido

AU - Martins, Isabelle

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AB - Imaging flow cytometry allows for the quantitative assessment of fluorescent signals at the subcellular level. Here, we describe the use of a biosensor cell line, namely, U2OS osteosarcoma cells equipped with a fusion protein comprising monomeric red fluorescent protein (mRFP), green fluorescent protein (GFP) and microtubule-associated proteins 1A/1B light chain 3B (best known as LC3), for the assessment of autophagic flux by imaging flow cytometry. We detail all analysis tools required to distinguish autophagosomes (that emit both a red and a green fluorescence) and autolysosomes (that emit a red fluorescence, yet lose the green fluorescent signal) and to quantitate autophagic flux in a convenient fashion.

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