内质网
细胞器
机械敏感通道
化学
生物物理学
膜
细胞生物学
内体
荧光显微镜
细胞内
线粒体
荧光
生物化学
小泡
生物
离子通道
量子力学
物理
受体
作者
Antoine Goujon,Adai Colom,Karolína Straková,Vincent Mercier,Dora Mahečić,Suliana Manley,Naomi Sakai,Aurélien Roux,Stefan Matile
摘要
Measuring forces inside cells is particularly challenging. With the development of quantitative microscopy, fluorophores which allow the measurement of forces became highly desirable. We have previously introduced a mechanosensitive flipper probe, which responds to the change of plasma membrane tension by changing its fluorescence lifetime and thus allows tension imaging by FLIM. Herein, we describe the design, synthesis, and evaluation of flipper probes that selectively label intracellular organelles, i.e., lysosomes, mitochondria, and the endoplasmic reticulum. The probes respond uniformly to osmotic shocks applied extracellularly, thus confirming sensitivity toward changes in membrane tension. At rest, different lifetimes found for different organelles relate to known differences in membrane organization rather than membrane tension and allow colabeling in the same cells. At the organelle scale, lifetime heterogeneity provides unprecedented insights on ER tubules and sheets, and nuclear membranes. Examples on endosomal trafficking or increase of tension at mitochondrial constriction sites outline the potential of intracellularly targeted fluorescent tension probes to address essential questions that were previously beyond reach.
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