Visualizing NBD-lipid Uptake in Mammalian Cells by Confocal Microscopy

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Visualizing NBD-lipid Uptake in Mammalian Cells by Confocal Microscopy. / Baum, Julia F.; Bredegaard, Lasse; Herrera, Sara Abad; Pomorski, Thomas Günther.

In: Bio-protocol, Vol. 13, No. 13, e4771, 2023.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Baum, JF, Bredegaard, L, Herrera, SA & Pomorski, TG 2023, 'Visualizing NBD-lipid Uptake in Mammalian Cells by Confocal Microscopy', Bio-protocol, vol. 13, no. 13, e4771. https://doi.org/10.21769/BioProtoc.4771

APA

Baum, J. F., Bredegaard, L., Herrera, S. A., & Pomorski, T. G. (2023). Visualizing NBD-lipid Uptake in Mammalian Cells by Confocal Microscopy. Bio-protocol, 13(13), [e4771]. https://doi.org/10.21769/BioProtoc.4771

Vancouver

Baum JF, Bredegaard L, Herrera SA, Pomorski TG. Visualizing NBD-lipid Uptake in Mammalian Cells by Confocal Microscopy. Bio-protocol. 2023;13(13). e4771. https://doi.org/10.21769/BioProtoc.4771

Author

Baum, Julia F. ; Bredegaard, Lasse ; Herrera, Sara Abad ; Pomorski, Thomas Günther. / Visualizing NBD-lipid Uptake in Mammalian Cells by Confocal Microscopy. In: Bio-protocol. 2023 ; Vol. 13, No. 13.

Bibtex

@article{9846f4c7e2cf4240be75d76f1c71d606,
title = "Visualizing NBD-lipid Uptake in Mammalian Cells by Confocal Microscopy",
abstract = "Eukaryotic cells use a series of membrane transporters to control the movement of lipids across their plasma membrane. Several tools and techniques have been developed to analyze the activity of these transporters in the plasma membrane of mammalian cells. Among them, assays based on fluorescence microscopy in combination with fluorescent lipid probes are particularly suitable, allowing visualization of lipid internalization in living cells. Here, we provide a step-by-step protocol for mammalian cell culture, lipid probe preparation, cell labeling, and confocal imaging to monitor lipid internalization by lipid flippases at the plasma membrane based on lipid probes carrying a fluorophore at a short-chain fatty acid. The protocol allows studying a wide range of mammalian cell lines, to test the impact of gene knockouts on lipid internalization at the plasma membrane and changes in lipid uptake during cell differentiation.",
keywords = "Confocal microscopy, Lipid transport, Mammalian cells, NBD-lipid, Plasma membrane",
author = "Baum, {Julia F.} and Lasse Bredegaard and Herrera, {Sara Abad} and Pomorski, {Thomas G{\"u}nther}",
note = "Publisher Copyright: {\textcopyright} 2023 The Authors.",
year = "2023",
doi = "10.21769/BioProtoc.4771",
language = "English",
volume = "13",
journal = "Bio-protocol",
issn = "2331-8325",
publisher = "bio-protocol",
number = "13",

}

RIS

TY - JOUR

T1 - Visualizing NBD-lipid Uptake in Mammalian Cells by Confocal Microscopy

AU - Baum, Julia F.

AU - Bredegaard, Lasse

AU - Herrera, Sara Abad

AU - Pomorski, Thomas Günther

N1 - Publisher Copyright: © 2023 The Authors.

PY - 2023

Y1 - 2023

N2 - Eukaryotic cells use a series of membrane transporters to control the movement of lipids across their plasma membrane. Several tools and techniques have been developed to analyze the activity of these transporters in the plasma membrane of mammalian cells. Among them, assays based on fluorescence microscopy in combination with fluorescent lipid probes are particularly suitable, allowing visualization of lipid internalization in living cells. Here, we provide a step-by-step protocol for mammalian cell culture, lipid probe preparation, cell labeling, and confocal imaging to monitor lipid internalization by lipid flippases at the plasma membrane based on lipid probes carrying a fluorophore at a short-chain fatty acid. The protocol allows studying a wide range of mammalian cell lines, to test the impact of gene knockouts on lipid internalization at the plasma membrane and changes in lipid uptake during cell differentiation.

AB - Eukaryotic cells use a series of membrane transporters to control the movement of lipids across their plasma membrane. Several tools and techniques have been developed to analyze the activity of these transporters in the plasma membrane of mammalian cells. Among them, assays based on fluorescence microscopy in combination with fluorescent lipid probes are particularly suitable, allowing visualization of lipid internalization in living cells. Here, we provide a step-by-step protocol for mammalian cell culture, lipid probe preparation, cell labeling, and confocal imaging to monitor lipid internalization by lipid flippases at the plasma membrane based on lipid probes carrying a fluorophore at a short-chain fatty acid. The protocol allows studying a wide range of mammalian cell lines, to test the impact of gene knockouts on lipid internalization at the plasma membrane and changes in lipid uptake during cell differentiation.

KW - Confocal microscopy

KW - Lipid transport

KW - Mammalian cells

KW - NBD-lipid

KW - Plasma membrane

U2 - 10.21769/BioProtoc.4771

DO - 10.21769/BioProtoc.4771

M3 - Journal article

C2 - 37456343

AN - SCOPUS:85164813598

VL - 13

JO - Bio-protocol

JF - Bio-protocol

SN - 2331-8325

IS - 13

M1 - e4771

ER -

ID: 382557690