Intravital imaging of fluorescent markers and FRET probes by DNA tattooing
Intravital imaging of fluorescent markers and FRET probes by DNA tattooing
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DOI：
10.1186/1472-6750-7-2
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发表时间：
2007-01-03
影响因子：
3.5
通讯作者：
Schumacher, Ton N. M.
中科院分区：
文献类型：
--
作者：
Bins, Adriaan D.;van Rheenen, Jacco;Schumacher, Ton N. M.
背景：荧光显微镜和小鼠转基因技术的进步使成像活体动物的分子事件成为可能。然而，转基因小鼠的产生是一个漫长的过程，活体成像需要专门的知识和设备。在这里，我们报道了一种使用通用设备的快速、非苛刻的活体成像方法。结果：通过DNA纹身，我们将编码荧光生物传感器的DNA转染活着的小鼠角质形成细胞。随后，表达这些生物传感器的单个细胞的行为可以在几个小时内使用传统的显微镜设备进行可视化。使用这种“瞬时转基因”模型，结合校正的坐标系，我们跟踪了几天来表达基于FRET或基于位置的生物传感器的单个细胞的体内行为。结论：这种“瞬时皮肤转基因”模型可以在纹身后的几天内跟踪表达FRET和定位探针的单个细胞的体内行为，为小鼠皮肤活体成像提供了一种快速、廉价的方法。
Background: Advances in fluorescence microscopy and mouse transgenesis have made it possible to image molecular events in living animals. However, the generation of transgenic mice is a lengthy process and intravital imaging requires specialized knowledge and equipment. Here, we report a rapid and undemanding intravital imaging method using generally available equipment.Results: By DNA tattooing we transfect keratinocytes of living mice with DNA encoding fluorescent biosensors. Subsequently, the behavior of individual cells expressing these biosensors can be visualized within hours and using conventional microscopy equipment. Using this "instant transgenic" model in combination with a corrected coordinate system, we followed the in vivo behavior of individual cells expressing either FRET- or location-based biosensors for several days. The utility of this approach was demonstrated by assessment of in vivo caspase-3 activation upon induction of apoptosis.Conclusion: This "instant skin transgenic" model can be used to follow the in vivo behavior of individual cells expressing either FRET- or location-based probes for several days after tattooing and provides a rapid and inexpensive method for intravital imaging in murine skin.