How To Characterize Individual Nanosize Liposomes with Simple Self-Calibrating Fluorescence Microscopy,Nano Letters

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How To Characterize Individual Nanosize Liposomes with Simple Self-Calibrating Fluorescence Microscopy
Nano Letters ( IF 9.6 ) Pub Date : 2018-04-03 00:00:00 , DOI: 10.1021/acs.nanolett.7b05312
Kim I. Mortensen ₁ , Chiara Tassone ₁ , Nicky Ehrlich ₁ , Thomas L. Andresen ₁ , Henrik Flyvbjerg ₁

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Nanosize lipid vesicles are used extensively at the interface between nanotechnology and biology, e.g., as containers for chemical reactions at minute concentrations and vehicles for targeted delivery of pharmaceuticals. Typically, vesicle samples are heterogeneous as regards vesicle size and structural properties. Consequently, vesicles must be characterized individually to ensure correct interpretation of experimental results. Here we do that using dual-color fluorescence labeling of vesicles—of their lipid bilayers and lumens, separately. A vesicle then images as two spots, one in each color channel. A simple image analysis determines the total intensity and width of each spot. These four data all depend on the vesicle radius in a simple manner for vesicles that are spherical, unilamellar, and optimal encapsulators of molecular cargo. This permits identification of such ideal vesicles. They in turn enable calibration of the dual-color fluorescence microscopy images they appear in. Since this calibration is not a separate experiment but an analysis of images of vesicles to be characterized, it eliminates the potential source of error that a separate calibration experiment would have been. Nonideal vesicles in the same images were characterized by how their four data violate the calibrated relationship established for ideal vesicles. In this way, our method yields size, shape, lamellarity, and encapsulation efficiency of each imaged vesicle. Applying this procedure to extruded samples of vesicles, we found that, contrary to common assumptions, only a fraction of vesicles are ideal.

中文翻译：

如何通过简单的自校准荧光显微镜表征单个纳米大小的脂质体

纳米级脂质囊泡被广泛用于纳米技术和生物学之间的界面，例如，用作微量浓度化学反应的容器和用于药物靶向输送的载体。通常，囊泡样品就囊泡大小和结构性质而言是异质的。因此，必须单独表征囊泡以确保正确解释实验结果。在这里，我们使用囊泡的双色荧光标记（分别是脂质双层和内腔）进行标记。然后，囊泡成像为两个斑点，每个颜色通道一个。简单的图像分析即可确定每个点的总强度和宽度。对于球形的，单层的和最佳的分子货物囊泡的囊泡，这四个数据都以简单的方式取决于囊泡半径。理想的囊泡。反过来，它们也可以对出现的双色荧光显微镜图像进行校准。由于此校准不是单独的实验，而是对要表征的囊泡图像的分析，因此消除了单独的校准实验可能存在的潜在误差来源。是。同一图像中的非理想囊泡的特征在于它们的四个数据如何违反为理想囊泡建立的校准关系。以这种方式，我们的方法产生每个成像囊泡的大小，形状，层状性和包封效率。将此程序应用于囊泡的挤压样品，我们发现，与通常的假设相反，只有一小部分囊泡是理想的。

更新日期：2018-04-03

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