Deborah J Rogers, Brennan P Bergeron, Glen M Watson, Karen Müller Smith
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引用次数: 0
Abstract
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 specialized 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 signaling. It was also used on slices obtained from 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.
活细胞成像是实验生物学的一项标准技术,可实时观察离体细胞和组织切片,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都需要使用专用显微镜和/或专门的平台适配器,而且成本相当高。我们采用三维打印技术制作了一种低成本的成像室,它带有用于交换液体的侧端口,可在直立显微镜上使用。该成像室提高了标准直立荧光显微镜的功能,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 刺激对钙信号转导的影响。它还可用于使用脑基质切片机获得的小鼠大脑切片。该样品室的优点包括:设计非常简单,可与直立式荧光显微镜配合使用,不需要任何特殊的平台适配器,并且包括可在成像过程中进行液体交换的端口。对于无法使用专用倒置荧光显微镜进行组织培养实验的本科生实验室来说,该室是理想的教育环境。
期刊介绍:
Journal of Biological Education is firmly established as the authoritative voice in the world of biological education.
The journal aims to bridge the gap between research and practice, providing information, ideas and opinion, in addition to critical examinations of advances in biology research and teaching. Through the coverage of policy and curriculum developments, the latest results of research into the teaching, learning and assessment of biology are brought to the fore.
Special emphasis is placed on research relevant to educational practice, guided by educational realities in systems, schools, colleges and universities. Papers that are theoretically informed and methodologically rigorous are welcomed.