一种基于荧光寿命的新方法,用于准确定量 BODIPY 活力保持的秀丽隐杆线虫的脂质。

IF 5 2区 医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY
Journal of Lipid Research Pub Date : 2024-10-01 Epub Date: 2024-09-19 DOI:10.1016/j.jlr.2024.100646
Chen Xu, Jintao Luo, Yong Yu
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引用次数: 0

摘要

脂滴(LDs)是与脂质储存和能量代谢相关的细胞器,因此其形态和数量具有重要的研究意义。市售的 BODIPY 染料能有效标记各种细胞类型中的脂滴,但它也能标记秀丽隐杆线虫的溶酶体相关细胞器(LRO),从而导致非特异性的脂滴定量。在这里,我们报告了 BODIPY 的荧光信号在 LROs 和 LDs 上表现出不同的荧光寿命模式,这可以通过荧光寿命成像显微镜来捕获、观察和过滤。此外,我们还提出并验证了一种基于荧光寿命的方法,该方法可提高 BODIPY 生命染色蠕虫脂肪储存定量的准确性,具有广泛的应用前景,包括在正向遗传筛选中快速准确地定量 LD。此外,我们的方法还能观察活体蠕虫中动态的 LD-LRO 相互作用,这是 BODIPY 生命染色的独特能力。我们的研究结果突显了 LD 和 LRO 不同的 BODIPY 荧光寿命特征,为未来研究 LD、LRO 或它们之间的相互作用提供了宝贵的工具。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
A fluorescence lifetime-based novel method for accurate lipid quantification of BODIPY vital-stained C. elegans.

Lipid droplets (LDs) are organelles associated with lipid storage and energy metabolism, thus, their morphology and quantity are of significant research interest. While commercially available BODIPY dye effectively labels LDs in various cell types, it also labels lysosome-related organelles (LROs) in C. elegans, leading to non-specific LD quantification. Here, we report that the fluorescent signals of BODIPY exhibit distinct fluorescence lifetime patterns for LROs and LDs, which can be captured, visualized, and filtered by fluorescence lifetime imaging microscopy. Furthermore, we proposed and validated a method based on fluorescence lifetime that can improve the accuracy of fat storage quantification in BODIPY vital-staining worms, which holds broad applications, including rapid and accurate LD quantification in forward genetic screening. Additionally, our method enables observing dynamic LD-LRO interactions in living worms, a unique capability of BODIPY vital staining. Our findings highlight distinct BODIPY fluorescence lifetime characteristics of LDs and LROs, providing a valuable tool for future research on LDs, LROs, or their interactions.

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来源期刊
Journal of Lipid Research
Journal of Lipid Research 生物-生化与分子生物学
CiteScore
11.10
自引率
4.60%
发文量
146
审稿时长
41 days
期刊介绍: The Journal of Lipid Research (JLR) publishes original articles and reviews in the broadly defined area of biological lipids. We encourage the submission of manuscripts relating to lipids, including those addressing problems in biochemistry, molecular biology, structural biology, cell biology, genetics, molecular medicine, clinical medicine and metabolism. Major criteria for acceptance of articles are new insights into mechanisms of lipid function and metabolism and/or genes regulating lipid metabolism along with sound primary experimental data. Interpretation of the data is the authors’ responsibility, and speculation should be labeled as such. Manuscripts that provide new ways of purifying, identifying and quantifying lipids are invited for the Methods section of the Journal. JLR encourages contributions from investigators in all countries, but articles must be submitted in clear and concise English.
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