Automated lipid droplet quantification system for phenotypic analysis of adipocytes using CellProfiler.
Automated lipid droplet quantification system for phenotypic analysis of adipocytes using CellProfiler.
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DOI：
10.1080/15376516.2020.1747124
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发表时间：
2020-06
影响因子：
3.2
通讯作者：
Veiga-Lopez A
中科院分区：
文献类型：
--
作者：
Adomshick V;Pu Y;Veiga-Lopez A
成脂分化是前脂肪细胞变成成熟脂肪细胞的过程，成熟脂肪细胞是储存能量和调节代谢稳态的细胞。在分化过程中，可以使用染色剂检测脂肪细胞中积累的中性脂质，并将其用作细胞分化的指标。然而，用于评估细胞内脂滴的成像工具仍处于起步阶段。营养、压力或化学物质暴露会导致脂肪形成分化和脂质代谢失调。因此，本研究的目的是开发一种准确、标准化的方法来量化成熟脂肪细胞的脂滴大小，并开发一种聚类方法来分析每个脂肪细胞的总脂质含量。对于脂滴分析，我们使用了两种方法，参考的免费在线计算机软件 ImageJ 和另一个免费在线计算机软件 CellProfiler。对于 ImageJ，我们使用了一个已经开发的宏，旨在识别粒子并量化其面积，对于 CellProfiler，我们开发了一个新的分析管道。我们的结果表明，与 ImageJ 相比，CellProfiler 能够准确识别更多数量的脂滴。还可以使用 CellProfiler 进行聚类分析，它可以量化每个脂肪细胞的总脂质含量，以深入了解单细胞对脂肪形成刺激的反应。与更传统的分析方法相比，CellProfiler 简化了脂肪细胞的脂滴表型分析。总之，这种新颖的图像分析工具可以对脂滴和脂肪生成失调进行更精确的评估，这是理解代谢紊乱的关键需求。
Adipogenic differentiation is the process by which preadipocytes become mature adipocytes, cells that store energy and regulate metabolic homeostasis. During differentiation, neutral lipids that accumulate in adipocytes can be detected using stains and used as an index of cell differentiation. However, imaging tools for evaluating intracellular lipid droplets remain at their infancy. Nutrition, stress, or chemical exposure can dysregulate adipogenic differentiation and lipid metabolism. Therefore, the aims of this study were to develop an accurate, standardized approach to quantify lipid droplet size of mature adipocytes and a clustering approach to analyze the total lipid content per adipocyte. For the lipid droplet analysis, we used two approaches, the free online computer software of reference, ImageJ, and another free online computer software, CellProfiler. For ImageJ, we used an already developed macro designed to identify particles and quantify their area, and for CellProfiler, we developed a new analysis pipeline. Our results show that CellProfiler is able to accurately identify a greater number of lipid droplets compared to ImageJ. A clustering analysis is also possible using CellProfiler which allows for the quantification of total lipid content per individual adipocyte to provide insight into single-cell responsiveness to adipogenic stimuli. CellProfiler streamlines the lipid droplet phenotypic analysis of adipocytes compared to more traditional analysis methods. In conclusion, this novel image analysis tool can provide with a more precise evaluation of lipid droplet and adipogenesis dysregulation, a critical need in the understanding of metabolic disorders.
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影响因子：
1.5
作者：
Al-Eryani L;Wahlang B;Falkner KC;Guardiola JJ;Clair HB;Prough RA;Cave M
通讯作者：
Cave M
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通讯作者：
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作者：
Exner, Tarik;Beretta, Carlo A.;Fuellekrug, Joachim
通讯作者：
Fuellekrug, Joachim