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Visualization of Lipid Droplets in the Alveolar Macrophage Cell Line MH-S with Live-cell Imaging by 3D Holotomographic Microscopy (Nanolive)

Lipid droplets (LD), triglycerides and sterol esters among them, are well known for their capacity as lipid storage organelles. Recently, they have emerged as critical cytoplasmic structures involved in numerous biological functions. LD storage is generated de novo by the cell and provides an energy...

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Autores principales: Pérez-Montero, Andrea, Zaragoza, Oscar, Luque, Alfonso, Hortelano, Sonsoles, Acebo, Paloma
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Bio-Protocol 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9993077/
https://www.ncbi.nlm.nih.gov/pubmed/36908642
http://dx.doi.org/10.21769/BioProtoc.4629
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author Pérez-Montero, Andrea
Zaragoza, Oscar
Luque, Alfonso
Hortelano, Sonsoles
Acebo, Paloma
author_facet Pérez-Montero, Andrea
Zaragoza, Oscar
Luque, Alfonso
Hortelano, Sonsoles
Acebo, Paloma
author_sort Pérez-Montero, Andrea
collection PubMed
description Lipid droplets (LD), triglycerides and sterol esters among them, are well known for their capacity as lipid storage organelles. Recently, they have emerged as critical cytoplasmic structures involved in numerous biological functions. LD storage is generated de novo by the cell and provides an energy reserve, lipid precursors, and cell protection. Moreover, LD accumulation can be observed in some pathologies as obesity, atherosclerosis, or lung diseases. Fluorescence imaging techniques are the most widely used techniques to visualize cellular compartments in live cells, including LD. Nevertheless, presence of fluorophores can damage subcellular components and induce cytotoxicity, or even alter the dynamics of the organelles. As an alternative to fluorescence microscopy, label-free techniques such as stimulated Raman scattering and coherent anti-stokes Raman scattering microscopy offer a solution to avoid the undesirable effects caused by dyes and fluorescent proteins, but are expensive and complex. Here, we describe a label-free method using live-cell imaging by 3D holotomographic microscopy (Nanolive) to visualize LD accumulation in the MH-S alveolar macrophage cell line after treatment with oleic acid, a monounsaturated fatty acid that promotes lipid accumulation.
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spelling pubmed-99930772023-03-09 Visualization of Lipid Droplets in the Alveolar Macrophage Cell Line MH-S with Live-cell Imaging by 3D Holotomographic Microscopy (Nanolive) Pérez-Montero, Andrea Zaragoza, Oscar Luque, Alfonso Hortelano, Sonsoles Acebo, Paloma Bio Protoc Methods Article Lipid droplets (LD), triglycerides and sterol esters among them, are well known for their capacity as lipid storage organelles. Recently, they have emerged as critical cytoplasmic structures involved in numerous biological functions. LD storage is generated de novo by the cell and provides an energy reserve, lipid precursors, and cell protection. Moreover, LD accumulation can be observed in some pathologies as obesity, atherosclerosis, or lung diseases. Fluorescence imaging techniques are the most widely used techniques to visualize cellular compartments in live cells, including LD. Nevertheless, presence of fluorophores can damage subcellular components and induce cytotoxicity, or even alter the dynamics of the organelles. As an alternative to fluorescence microscopy, label-free techniques such as stimulated Raman scattering and coherent anti-stokes Raman scattering microscopy offer a solution to avoid the undesirable effects caused by dyes and fluorescent proteins, but are expensive and complex. Here, we describe a label-free method using live-cell imaging by 3D holotomographic microscopy (Nanolive) to visualize LD accumulation in the MH-S alveolar macrophage cell line after treatment with oleic acid, a monounsaturated fatty acid that promotes lipid accumulation. Bio-Protocol 2023-03-05 /pmc/articles/PMC9993077/ /pubmed/36908642 http://dx.doi.org/10.21769/BioProtoc.4629 Text en Copyright © 2023 The Authors; exclusive licensee Bio-protocol LLC. https://creativecommons.org/licenses/by-nc/4.0/This is an open access article under the CC BY-NC license (https://creativecommons.org/licenses/by-nc/4.0/).
spellingShingle Methods Article
Pérez-Montero, Andrea
Zaragoza, Oscar
Luque, Alfonso
Hortelano, Sonsoles
Acebo, Paloma
Visualization of Lipid Droplets in the Alveolar Macrophage Cell Line MH-S with Live-cell Imaging by 3D Holotomographic Microscopy (Nanolive)
title Visualization of Lipid Droplets in the Alveolar Macrophage Cell Line MH-S with Live-cell Imaging by 3D Holotomographic Microscopy (Nanolive)
title_full Visualization of Lipid Droplets in the Alveolar Macrophage Cell Line MH-S with Live-cell Imaging by 3D Holotomographic Microscopy (Nanolive)
title_fullStr Visualization of Lipid Droplets in the Alveolar Macrophage Cell Line MH-S with Live-cell Imaging by 3D Holotomographic Microscopy (Nanolive)
title_full_unstemmed Visualization of Lipid Droplets in the Alveolar Macrophage Cell Line MH-S with Live-cell Imaging by 3D Holotomographic Microscopy (Nanolive)
title_short Visualization of Lipid Droplets in the Alveolar Macrophage Cell Line MH-S with Live-cell Imaging by 3D Holotomographic Microscopy (Nanolive)
title_sort visualization of lipid droplets in the alveolar macrophage cell line mh-s with live-cell imaging by 3d holotomographic microscopy (nanolive)
topic Methods Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9993077/
https://www.ncbi.nlm.nih.gov/pubmed/36908642
http://dx.doi.org/10.21769/BioProtoc.4629
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