EVOS™ 60X 物镜，萤石，LWD

当在Z轴（上和下）聚焦得太高时，物镜会擦到容器支架适配器。这是EVOS FL Auto成像系统启动、仪器启动时移动载物台或切换物镜时遇到的特殊问题。盖玻片矫正物镜的镜筒顶部更加宽和平，尤其是在样品容器边缘成像时，这意味着它们更容易触碰容器支架适配器的边缘，在这些情况下，无法使用物镜在这些区域成像。如果物镜被容器支架适配器‘’卡‘’住了，慢慢的旋转容器支架适配器上的拇指螺丝，将其竖直提起离开载物台，然后再朝着载物台的中心向下移动物镜聚焦。最好的办法是在您的实验室设定一个关机流程，包括将物镜移动到最低的放大倍数，在每天关闭仪器之前向下调整镜头。

物镜镜头在与容器支架适配器的刮擦时会严重损毁。如果发生刮擦，取出物镜，仔细检查物镜——尤其是镜头——是否损坏。 

如果镜头一直在样品上移动，可能是对焦太快且丢失焦平面（如果是手动聚焦）的问题，或者是物镜校准的问题（如果使用自动聚焦）。最好使用系统自带的FL Auto校准载玻片校准物镜。首先检查看您的物镜是长工作距离（LWD）物镜或盖玻片矫正（CC）物镜。如果是盖玻片矫正（CC）物镜，则只能采用超短的工作距离，通过薄盖玻片进行成像，不能通过载玻片、微孔板或培养皿内的塑料底进行成像。如果使用高放大倍数或者油镜，利用肉眼观察，向上移动物镜接触到样品的底部，然后缓慢地朝远离样品的方向移动物镜，进一步聚焦。 物镜镜头在与样品的刮擦中会受到严重的损毁。如果发生刮擦，检查物镜的损坏情况。

•当颜色和焦距已被标准校准时，对于一般的使用，Plan Achromat物镜是绝佳的；这些适用于放大倍数要求低（2x到4x）的样品。推荐Plan Achromat物镜用于基本视野明亮的显微镜和简单荧光检测。
•Plan Fluorite物镜为明亮的荧光信号和高对比度提供了更高水平的分辨率。推荐基本的荧光成像以及更大放大倍数的明场显微镜使用这些物镜。
•Plan Apochromat物镜提供了最高水平的分辨率、荧光亮度、对比度和色度校准。如果您需要成像非常小的结构且需要高对比度和亮度，Plan Apochromat物镜是最好的选择。

所有的EVOS成像系统都是倒置显微镜。对CC物镜来说，盖玻片必须朝下，朝向物镜，因为镜头有一个仅适用于薄玻璃盖玻片或塑料盖玻片的短工作距离。LWD物镜设计用于观察微孔板、皮氏培养皿或者细胞培养瓶底部；这些物镜镜头的长工作距离适应于较厚的材料，诸如多种容器的塑料底。 

Objectives can hit the vessel holder when they are focused too high in the Z axis (up and down). This is a particularly a problem with the EVOS FL Auto Imaging System during instrument start-up, when the stage moves during system initiation, or when changing objectives. Coverslip-corrected objectives tend to be wider and flatter at the top of the barrel, which means that they are more likely to run into the edges of the vessel holder, particularly if you are imaging at the edges of the sample container. In those cases, use of that objective for those areas of the container may not be possible. If the objective if “jammed” by the vessel holder, then carefully unscrew the thumbscrews of the vessel holder and lift it straight off the stage, then move the objective downward in focus and toward the center of the stage. It is a good idea to have a shut-down procedure in your lab that includes moving the objectives to the lowest magnification and focusing downward with course focus prior to turning off the instrument for the day.
An objective can be damaged by scraping against the vessel holder. If this happens, take out the objective and examine it carefully for damage, particularly on the lens.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.