Our group is looking into getting a Countess II FL Cell Counter from Life Technologies (link). If anyone has had experience with this machine we would love to get some reviews to help with our decision. Thanks!
http://www.lifetechnologies.com/us/en/home/brands/product-brand/countess-automated-cell-counter.html
Hi Karl,
did you in the end decide to buy the Countess II FL and if yes what are your experiences with it? Thanks a lot for sharing your experiences!
Best,
Gernot
Hello. We did buy it but ended up returning it to the firm, because it didn't work well for what we needed. The Countess II works well for homogeneous cell preparations (ie cell lines), but it doesn't do really well with heterogeneous preparations (say whole spleen cells, whole peritoneal cells, etc.). We now bought a Nexcelom Cellometer Auto 2000 (more expensive), which is working much better for the heterogeneous cell preparations.
Thanks Alvaro for the comment as we have been testing a demo Countess II today in our lab and found that it would not count 5-10 um mouse lymphocytes. The instrument tags the cells as 80-90% dead with Trypan Blue staining on the brightfield default settings for the instrument and adjusting the gating does not seem to help. At first we thought we had bad cell preps but we confirmed our techniques via old fashioned hemacytometer counts. Clearly this instrument has some resolution problems for murine cell types. We will try further adjustments on the gating to see if we can overcome our initial problems. Maybe we are missing something with the gating strategy for murine lymphocytes.
Thanks for the comment, Alan. It is reassuring that people in other labs encounter the same problems.
Trypan blue is not a good method for counting small cells like PBMC or other lymphocytes. For both people and instruments its hard to tell if a cell is dark because it is small and out of focus or it is indeed TB positive. It is best to use Fluorescent methods that use nuclear stains like acridine orange and propidum iodide (like on the auto 2000 that Alvaro has). The added benefit here is that the nuclear stains do not stain debris, so you end up with a cell count that does not include RBC or debris.
The countess II should be able to do fluorescence if you get the cubes... I have however never seen one in a lab.. they all seem to be the black and white ones.
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