EVOS™ FL Color Imaging System

With the EVOS FL Cell Imaging System (and the EVOS FL Color Imaging System), there is a ''live'' mode which shows the field of view live while you orient your sample. When you are imaging in ''live'' mode, the system increases the gain setting so it can have a quicker refresh rate. (Note: The EVOS FL doesn't allow manual adjustments of the gain setting). This makes it easier to move around the sample to orient yourself and see your sample without having long pauses due to longer exposure time. This also helps to reduce photobleaching. With a higher gain, you have a lower exposure time (but you also have higher background).

You will then capture the image with either ''Find and Focus'' mode or ''Actual'' mode. If using ''Find and Focus'' mode, when you capture the final image, the system will reset the exposure time and gain setting to optimize the contrast for your sample, which may be different than what you see in ''live'' mode. You can only adjust the light intensity in ''Find and Focus'' mode. ''Find and Focus'' mode is often best for qualitative imaging. If this is not conducive for your assay, or if you are performing quantitative assays that require more normalization across samples, then you will want to use ''Actual'' mode where you can control the exposure time yourself as well as the light intensity.

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

For the EVOS imaging systems:

Make certain the light is on (easy way to test this: place a piece of thin paper on the stage).
Make certain the sample is not too opaque; compare with a calibration slide or another, thinner or single-cell sample on a slide.
Check the objectives to make certain the turret is in alignment and the objective is completely threaded in its slot.
For the EVOS FL Imaging System: change the position of the light cubes.
For the EVOS FL Auto Imaging System: check the insertion of all USB ports for connectively from scope to computer.
For brightfield settings, check the condenser slider slot; make certain the condenser sliders are completely in place.

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

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.

Here is a page you can go to - https://www.thermofisher.com/us/en/home/products-and-services/services/instrument-qualification-services/instruments-and-services-portal/instrument-software-downloads.html. Look under the “Cell Imaging Systems” section and follow the link for your EVOS imaging system. There you will find the free download link as well as instructions to follow. We recommend you check for updates at least every six months, or if your system seems to have a software glitch of any sort.

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

Only the EVOS FL Auto Imaging System and EVOS FL Auto 2 Imaging System have an automatic cell counting function. On the EVOS FL Auto Imaging System, automatic cell counting is enabled via built-in instrument firmware whereas on the EVOS FL Auto 2 Imaging System, the Celleste Image Analysis Software needs to be purchased separately to enable automatic cell counting.

The EVOS XL, EVOS FL, and EVOS FL Color imaging systems provide a manual cell counting tool that allows tagging of up to six different labels on the screen image.

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