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There are a multitude of culture and imaging chambers available for cell imaging, but choosing the right one relies on good planning of your experiment before you start. Are you performing multiple treatments? Do the cells require special temperature, atmospheric, or light conditions? What's your microscope configuration?
The information below will help you understand the planning that you need to carry out to ensure that your cells are cultured and contained in a vessel that will allow you to capture the images you need.
There are several types of vessels through which you can image. Which one you choose depends on the type of experiment you are doing and the capabilities of your microscope.
The type of vessel you use will also depend on your microscope configuration. Here are some of the configuration details you’ll need to consider:
The type of vessel a microscope can accommodate will partly depend on whether the objectives are below (inverted microscope) or above (upright microscope) the stage (Figure 1).
An upright microscope is good for imaging samples on glass coverslips. This is because the objective can get very close to the coverslip containing the sample. An upright microscope, however is not the best choice for culture dishes and flasks. In these vessels, the cells you want to image are at the bottom of the dish, and a top-mounted objective would have to image through all of the air between it and the sample. Most objectives won’t be able to do this with good results.
Figure 1. The position of your microscope’s objective relative to the sample stage will determine which kinds of vessels will work the best for your imaging experiment.
Some stages require an adaptor to fit certain types of vessels. Double check that the microscope you will use has the configuration, holders, or adaptor accessories you need to hold your vessel in place while imaging.
You will need to use an objective with a working distance that matches the vessel your sample is in (Figure 2).
Sometimes the design of your experiment will limit the vessel type you can use. Here are some examples:
Figure 2. Upright microscopes generally work best with samples that are on glass coverslips or slides.
请在实验开始前确定细胞的存放方式尽管有很多培养和成像的器皿可供细胞成像实验使用,但能否选择到最适宜的器皿仍依赖于实验开始前周密的计划。是否要进行多重处理?细胞是否需要特定的温度、空气或光照条件?显微镜是什么配置? 下面提供的信息可帮助您了解应该如何进行实验规划,才能确保细胞在培养和存放的实验器皿中能获得所需的图像。 |
可供成像实验使用的器皿有多种类型,具体选择哪一种,取决于所进行的实验类型以及您所使用的显微镜的性能。
玻璃底器皿 | 多孔板 | 培养瓶 | ||
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您所选的器皿类型还取决于您的显微镜配置,以下为您需要考虑到的一些配置信息:
您使用的是正置显微镜还是倒置显微镜?微镜兼容的器皿类型部分取决于物镜处于载物台下方(倒置显微镜)还是上方 (正置显微镜)(图1). 正置显微镜适于对玻璃盖玻片上的样品成像,这是因为物镜可以非常接近盛放样品的盖玻片。但在使用培养皿和培养瓶时,正置显微镜就不是最好的选择,因为在这些器皿中,成像的细胞处于器皿的底部,而装在顶部的物镜必须透过样品和物镜之间的全部空气才能成像,大多数正置物镜都无法得到理想的效果。 图 1. 显微镜物镜相对于样品载物台的位置将决定哪种器皿最适合于您的成像实验。 |
有些载物台需要接适配器才能适用某些类型的器皿,请务必确认自己将要使用的显微镜,确认其具有适合在成像时固定您的实验器皿的配置、支架或适配器附件。
您要使用哪种类型的物镜?您需要使用匹配物镜工作距离的样品器皿 (图 2). 是否存在一些实验设计方面的注意事项?有时,您的实验设计会限制您所能使用的实验器皿类型,以下为部分实例:
图 2. 对于在玻璃盖玻片或载片上的样品,通常正置显微镜最适合。 |
仅供科研使用,不可用于诊断目的。