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首页> 外文期刊>Biophysical Journal >Label-free imaging of lipid-droplet intracellular motion in early Drosophila embryos using femtosecond-stimulated Raman loss microscopy
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Label-free imaging of lipid-droplet intracellular motion in early Drosophila embryos using femtosecond-stimulated Raman loss microscopy

机译：使用飞秒刺激的拉曼损失显微镜对果蝇早期胚胎中脂质滴的细胞内运动进行无标记成像

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摘要

Lipid droplets are complex organelles that exhibit highly dynamic behavior in early Drosophila embryo development. Imaging lipid droplet motion provides a robust platform for the investigation of shuttling by kinesin and dynein motors, but methods for imaging are either destructive or deficient in resolution and penetration to study large populations of droplets in an individual embryo. Here we report real-time imaging and quantification of droplet motion in live embryos using a recently developed technique termed "femtosecond-stimulated Raman loss" microscopy. We captured long-duration time-lapse images of the developing embryo, tracked single droplet motion within large populations of droplets, and measured the velocity and turning frequency of each particle at different apical-to-basal depths and stages of development. To determine whether the quantities for speed and turning rate measured for individual droplets are sufficient to predict the population distributions of droplet density, we simulated droplet motion using a velocity-jump model. This model yielded droplet density distributions that agreed well with experimental observations without any model optimization or unknown parameter estimation, demonstrating the sufficiency of a velocity-jump process for droplet trafficking dynamics in blastoderm embryos.
机译：脂质液滴是复杂的细胞器，在果蝇早期胚胎发育中表现出高度动态的行为。成像脂滴运动为驱动蛋白和动力蛋白运动的穿梭研究提供了一个强大的平台，但是成像方法对研究单个胚胎中的大量液滴而言是破坏性的或分辨率和穿透性不足。在这里，我们报告了实时成像，并使用一种称为“飞秒刺激的拉曼损失”显微镜的最新技术对活体胚​​胎中的液滴运动进行了量化。我们捕获了发育中的胚胎的长时间延时图像，跟踪了大液滴群中的单个液滴运动，并测量了从不同的根尖到基部深度和发育阶段的每个粒子的速度和转向频率。为了确定单个液滴的速度和转动速率量是否足以预测液滴密度的总体分布，我们使用速度跳跃模型模拟了液滴运动。该模型产生的液滴密度分布与实验观察结果非常吻合，而没有任何模型优化或未知参数估计，证明了胚盘胚中液滴运输动态的速度跳跃过程是足够的。

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