Lipid droplet quantification based on iterative image processing[S]
Lipid droplets (LDs) are ubiquitous and highly dynamic subcellular organelles required for the storage of neutral lipids. LD number and size distribution are key parameters affected not only by nutrient supply but also by lipotoxic stress and metabolic regulation. Current methods for LD quantificati...
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doaj-f5006bee3dbd4c18b2319d5e0b6adb5d2021-04-29T04:34:53ZengElsevierJournal of Lipid Research0022-22752019-07-0160713331344Lipid droplet quantification based on iterative image processing[S]Tarik Exner0Carlo A. Beretta1Qi Gao2Cassian Afting3Inés Romero-Brey4Ralf Bartenschlager5Leonard Fehring6Margarete Poppelreuther7Joachim Füllekrug8To whom correspondence should be addressed; Molecular Cell Biology Laboratory Internal Medicine IV,Heidelberg University, Heidelberg, Germany; To whom correspondence should be addressedCellNetworks Math-Clinic Core Facility, BioQuantHeidelberg University, Heidelberg, GermanyCellNetworks Math-Clinic Core Facility, BioQuantHeidelberg University, Heidelberg, GermanyMolecular Cell Biology Laboratory Internal Medicine IV,Heidelberg University, Heidelberg, GermanyDepartment of Infectious Diseases, Molecular VirologyHeidelberg University, Heidelberg, GermanyDepartment of Infectious Diseases, Molecular VirologyHeidelberg University, Heidelberg, Germany; Department of Virus-Associated Carcinogenesis,German Cancer Research Center, Heidelberg, GermanyMolecular Cell Biology Laboratory Internal Medicine IV,Heidelberg University, Heidelberg, GermanyMolecular Cell Biology Laboratory Internal Medicine IV,Heidelberg University, Heidelberg, GermanyMolecular Cell Biology Laboratory Internal Medicine IV,Heidelberg University, Heidelberg, GermanyLipid droplets (LDs) are ubiquitous and highly dynamic subcellular organelles required for the storage of neutral lipids. LD number and size distribution are key parameters affected not only by nutrient supply but also by lipotoxic stress and metabolic regulation. Current methods for LD quantification lack general applicability and are either based on time consuming manual evaluation or show limitations if LDs are high in numbers or closely clustered. Here, we present an ImageJ-based approach for the detection and quantification of LDs stained by neutral lipid dyes in images acquired by conventional wide-field fluorescence microscopy. The method features an adjustable preprocessing procedure that resolves LD clusters. LD identification is based on their circular edges and central fluorescence intensity maxima. Adaptation to different cell types is mediated by a set of interactive parameters. Validation was done for three different cell lines using manual evaluation of LD numbers and volume measurement by 3D rendering of confocal datasets. In an application example, we show that overexpression of the acyl-CoA synthetase, FATP4/ACSVL5, in oleate-treated COS7 cells increased the size of LDs but not their number.http://www.sciencedirect.com/science/article/pii/S0022227520310646endoplasmic reticulumfatty acid/metabolismfluorescence microscopytriglyceridesimage quantificationImageJ/Fiji |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Tarik Exner Carlo A. Beretta Qi Gao Cassian Afting Inés Romero-Brey Ralf Bartenschlager Leonard Fehring Margarete Poppelreuther Joachim Füllekrug |
spellingShingle |
Tarik Exner Carlo A. Beretta Qi Gao Cassian Afting Inés Romero-Brey Ralf Bartenschlager Leonard Fehring Margarete Poppelreuther Joachim Füllekrug Lipid droplet quantification based on iterative image processing[S] Journal of Lipid Research endoplasmic reticulum fatty acid/metabolism fluorescence microscopy triglycerides image quantification ImageJ/Fiji |
author_facet |
Tarik Exner Carlo A. Beretta Qi Gao Cassian Afting Inés Romero-Brey Ralf Bartenschlager Leonard Fehring Margarete Poppelreuther Joachim Füllekrug |
author_sort |
Tarik Exner |
title |
Lipid droplet quantification based on iterative image processing[S] |
title_short |
Lipid droplet quantification based on iterative image processing[S] |
title_full |
Lipid droplet quantification based on iterative image processing[S] |
title_fullStr |
Lipid droplet quantification based on iterative image processing[S] |
title_full_unstemmed |
Lipid droplet quantification based on iterative image processing[S] |
title_sort |
lipid droplet quantification based on iterative image processing[s] |
publisher |
Elsevier |
series |
Journal of Lipid Research |
issn |
0022-2275 |
publishDate |
2019-07-01 |
description |
Lipid droplets (LDs) are ubiquitous and highly dynamic subcellular organelles required for the storage of neutral lipids. LD number and size distribution are key parameters affected not only by nutrient supply but also by lipotoxic stress and metabolic regulation. Current methods for LD quantification lack general applicability and are either based on time consuming manual evaluation or show limitations if LDs are high in numbers or closely clustered. Here, we present an ImageJ-based approach for the detection and quantification of LDs stained by neutral lipid dyes in images acquired by conventional wide-field fluorescence microscopy. The method features an adjustable preprocessing procedure that resolves LD clusters. LD identification is based on their circular edges and central fluorescence intensity maxima. Adaptation to different cell types is mediated by a set of interactive parameters. Validation was done for three different cell lines using manual evaluation of LD numbers and volume measurement by 3D rendering of confocal datasets. In an application example, we show that overexpression of the acyl-CoA synthetase, FATP4/ACSVL5, in oleate-treated COS7 cells increased the size of LDs but not their number. |
topic |
endoplasmic reticulum fatty acid/metabolism fluorescence microscopy triglycerides image quantification ImageJ/Fiji |
url |
http://www.sciencedirect.com/science/article/pii/S0022227520310646 |
work_keys_str_mv |
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