Distinguishing modes of cell death using the ImageStream® multispectral imaging flow cytometer
Distinguishing modes of cell death using the ImageStream® multispectral imaging flow cytometer
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DOI：
10.1002/cyto.a.20048
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发表时间：
2004-06-01
期刊：
影响因子：
3.7
通讯作者：
Morrissey, PJ
中科院分区：
文献类型：
--
作者：
George, TC;Basiji, DA;Morrissey, PJ
背景：在这里，我们展示了ImageStream(R) 100多光谱成像细胞仪区分活细胞、坏死细胞、早期和晚期凋亡细胞的能力，使用独特的光度和形态特征组合。方法：制备活细胞、坏死细胞、早期和晚期凋亡细胞，并通过免疫荧光显微镜、常规流式细胞术和成像流式细胞术，分别作为单个群体和异质细胞混合物进行分析。结果：活细胞（膜联蛋白V-， 7-AAD(-)）和早期凋亡细胞（膜联蛋白V+， 7-AAD(-)）均可通过常规或基于ImageStream的流式细胞术检测。然而，对细胞的多光谱图像检查表明，膜联蛋白V+， 7-AAD（+）群体包含坏死和晚期凋亡细胞。虽然这些细胞不能用传统的流式细胞技术区分，但它们可以用ImageStream技术提供的光度和形态测量的独特组合来分离。结论：使用多光谱图像，可以使用传统流式细胞仪无法提供的特征来区分形态学上不同的细胞群。特别是，形态测量与光度测量相结合的能力使得区分活细胞与凋亡早期细胞，以及晚期凋亡细胞与坏死细胞成为可能。(C) 2004 Wilcy-I,iss, Inc。
Background: Here we demonstrate the ability of the, ImageStream(R) 100 Multispectral Imaging Cytometer to discriminate between live, necrotic, and early and late apoptotic cells, using unique combinations of photometric and morphometric features.Methods: Live, necrotic, and early and late apoptotic cells were prepared and analyzed by immunofluorescence microscopy, conventional flow Cytotuctry, and imaging flow cytometry, both as single populations and as a heterogeneous mixture of cells.Results: Live (annexin V-, 7-AAD(-)) and early apoptotic (annexin V+, 7-AAD(-)) cells were readily identifiable using either conventional or ImageStream based flow cytometric methods. However, inspection of multispectral images of cells demonstrated that the annexin V+, 7-AAD(+) population contained both necrotic and late-stage apoptotic cells. Although these cells could not be distinguished using conventional flow cytometric techniques, they were separable using unique combinations of photometric and morphometric measures available using ImageStream technologies.Conclusions: Using multispectral imagery, morphologically distinct cell populations can be distinguished using features not available with conventional flow cytometers. In particular, the ability to couple morphometric with photometric measures makes it possible to distinguish live cells from cells in the early phases of apoptosis, as well as late apoptotic cells from necrotic cells. (C) 2004 Wilcy-I,iss, Inc.