3D printed chamber for live cell imaging on an upright epifluorescence microscope.
3D printed chamber for live cell imaging on an upright epifluorescence microscope.
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用于在直立落射荧光显微镜上进行活细胞成像的 3D 打印室。
DOI：
10.1080/00219266.2022.2030389
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发表时间：
2024
影响因子：
1.1
通讯作者：
Smith,KarenMüller
中科院分区：
文献类型：
--
作者：
Rogers,DeborahJ;Bergeron,BrennanP;Watson,GlenM;Smith,KarenMüller
活细胞成像是实验生物学中的一种标准技术，可以实时观察分离的细胞和组织切片并测试细胞对缓冲液成分变化的反应。然而，大多数活细胞成像设备需要使用专用显微镜和/或专用载物台适配器，并且成本相当高。我们采用 3D 打印技术创建了一个低成本成像室，该成像室带有用于交换液体的侧端口，可用于正置显微镜。该室增强了标准直立式落射荧光显微镜的功能，可对培养的小鼠下丘脑星形胶质细胞进行动态、实时钙成像，并测试 ATP 刺激对钙信号传导的影响。它还用于使用脑基质切片机从小鼠大脑获得的切片。该腔室的优点包括非常简单的设计，可与直立式落射荧光显微镜一起使用，不需要任何特殊的载物台适配器，并且包括允许在成像期间进行流体交换的端口。该室非常适合本科生实验室的教育环境，这些实验室无法使用专用的倒置荧光显微镜进行组织培养实验。
Live cell imaging is a standard technique in experimental biology that enables the observation of isolated cells and tissue slices in real time and the testing of cellular responses to changes in buffer composition. However, most live cell imaging devices require the use of dedicated microscopes and/or specialised stage adaptors, and come at a reasonably high cost. We employed 3D printing technology to create a low-cost imaging chamber with side ports to exchange fluids, to be used on upright microscopes. The chamber increased the functionality of a standard upright epifluorescent microscope to allow dynamic, real-time calcium imaging of cultured hypothalamic astrocytes from mice, and to test the effects of ATP stimulation upon calcium signalling. It was also used on slices obtained from the mouse brain using a brain matrix slicer. The advantages of this chamber include a very simple design that can be used with upright epifluorescence microscopes, does not require any special stage adaptor, and includes ports to permit fluid exchange during imaging. This chamber is ideal for educational settings with undergraduate laboratories that do not have access to dedicated inverted fluorescent microscopes for tissue culture experiments.