Frigault Melanie M, Lacoste Judith, Swift Jody L, Brown Claire M
Molecular Oncology Group, McGill University Cancer Centre, Montreal, Canada.
J Cell Sci. 2009 Mar 15;122(Pt 6):753-67. doi: 10.1242/jcs.033837.
Imaging of living cells and tissue is now common in many fields of the life and physical sciences, and is instrumental in revealing a great deal about cellular dynamics and function. It is crucial when performing such experiments that cell viability is at the forefront of any measurement to ensure that the physiological and biological processes that are under investigation are not altered in any way. Many cells and tissues are not normally exposed to light during their life cycle, so it is important for microscopy applications to minimize light exposure, which can cause phototoxicity. To ensure minimal light exposure, it is crucial that microscope systems are optimized to collect as much light as possible. This can be achieved using superior-quality optical components and state-of-the-art detectors. This Commentary discusses how to set up a suitable environment on the microscope stage to maintain living cells. There is also a focus on general and imaging-platform-specific ways to optimize the efficiency of light throughput and detection. With an efficient optical microscope and a good detector, the light exposure can be minimized during live-cell imaging, thus minimizing phototoxicity and maintaining cell viability. Brief suggestions for useful microscope accessories as well as available fluorescence tools are also presented. Finally, a flow chart is provided to assist readers in choosing the appropriate imaging platform for their experimental systems.
活细胞和组织成像如今在生命科学和物理科学的许多领域都很常见,有助于揭示大量有关细胞动力学和功能的信息。在进行此类实验时,至关重要的是,细胞活力应处于任何测量的首要位置,以确保所研究的生理和生物学过程不会以任何方式改变。许多细胞和组织在其生命周期中通常不会暴露于光线下,因此对于显微镜应用来说,尽量减少可能导致光毒性的光照至关重要。为确保光照最少,优化显微镜系统以尽可能多地收集光线至关重要。这可以通过使用优质光学组件和最先进的探测器来实现。本评论讨论了如何在显微镜载物台上设置合适的环境以维持活细胞。还重点介绍了优化光通量和检测效率的通用方法以及特定成像平台的方法。借助高效的光学显微镜和良好的探测器,在活细胞成像过程中可以将光照降至最低,从而将光毒性降至最低并维持细胞活力。还介绍了有用的显微镜附件以及可用荧光工具的简要建议。最后,提供了一个流程图,以帮助读者为其实验系统选择合适的成像平台。