关键词： AFM CP: Biotechnology CP: Cell biology FCS FLIM FRAP atomic force microscopy diffusion fluorescence correlation spectroscopy fluorescence lifetime imaging microscopy fluorescence microscopy fluorescence recovery after photobleaching model lipid droplets pendant droplet tensiometry phospholipid monolayers sorting tension

Mesh ： Lipid Droplets / metabolism chemistry Microscopy, Atomic Force / methods Microscopy, Fluorescence / methods Fluorescence Recovery After Photobleaching / methods Humans Flow Cytometry / methods Spectrometry, Fluorescence / methods

来  源：   DOI：10.1016/j.crmeth.2024.100774   PDF(Pubmed)

Abstract：
We present methods for making and testing the membrane biophysics of model lipid droplets (LDs). Methods are described for imaging LDs ranging in size from 0.1 to 40 μm in diameter with high-resolution microscopy and spectroscopy. With known LD compositions, membrane binding, sorting, diffusion, and tension were measured via fluorescence correlation spectroscopy (FCS), fluorescence recovery after photobleaching (FRAP), fluorescence lifetime imaging microscopy (FLIM), atomic force microscopy (AFM), and imaging flow cytometry. Additionally, a custom, small-volume pendant droplet tensiometer is described and used to measure the association of phospholipids to the LD surface. These complementary, cross-validating methods of measuring LD membrane behavior reveal the interplay of biophysical processes on lipid droplet monolayers.
摘要：
我们介绍了制造和测试模型脂滴（LD）的膜生物物理学的方法。描述了使用高分辨率显微镜和光谱学对直径为0.1至40μm的LD进行成像的方法。使用已知的LD组合物，膜结合，排序，扩散,和张力通过荧光相关光谱(FCS)测量，光漂白后的荧光恢复（FRAP），荧光寿命成像显微镜（FLIM），原子力显微镜（AFM），和成像流式细胞术。此外,一个习俗,描述了小体积悬垂液滴张力计，并用于测量磷脂与LD表面的缔合。这些互补的，测量LD膜行为的交叉验证方法揭示了脂滴单层上生物物理过程的相互作用。
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