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Knowledge

What to look for in an automated cell counter?

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Making the move: How to go digital

Why go digital when manual works, you may think? You learned it from your colleague who worked at the lab for decades. Sure he knows what the best practices are.

In summary?
  1. Why consider to go digital?
  2. Accurate cell counting focusing.
  3. Reproduction of results
  4. Cell counting specificity.
  5. Declustering performance with the automated cell counters .

Making the switch

Why consider to go digital?

Hemocytometer or the Bürker-Türk chamber was the standard tool decades ago. These techniques show some disadvantages as they are very time-consuming and error-prone. In the era of smart TVs and the internet carried in your pocket, I bet you prefer to use a fast, accurate, reproducible, and automated technique for cell counting, too. There are several automated cell counters on the market. But do you know what to look for in those systems?

Here are some tips for knowing what to look for in an automated cell counter.
Hemocytometer for cell counting

Accuracy

Accurate cell counter focusing

When preparing your cell suspension, it is difficult to get completely rid of dead cells or debris. You don't want to count any of those as a living cell. Your cell counter can only exclude dead cells and debris if it recognizes these particles. Therefore, accurate focusing is crucial. Automated cell counters detect living cells with black edges and white centers. Inaccurate focusing will mess up the black and white pixels, leading to misinterpretation of dead and living cells. Make sure your choice of cell counter is up to the task by checking its camera resolution. You can also choose a counter with autofocusing technology to keep things simple.

Reproducibility

Reproduction of results

Like all types of experiments, you also want your cell counting to be reproducible. Most automated cell counters provide you with specific slides that have a defined measuring area. Therefore, the sizes of the taken images are always equal for each count, eliminating variation in the field of view.
Image of cell with Luna II

Count all cells

Declustering performance with the automated cell counters

Although you do the best you can, your cell suspensions can still contain clustered cells after preparation. You want these cell groups to be counted as individual cells to get a fair cell count. Most systems have integrated software that automatically recognizes cell clusters and count the cells in these groups as individual cells. But don't think about it too quickly; the declustering performance varies significantly between the various available cell counters!

So when you take a little time to get yourself a decent comparison of automated cell counters, be sure to include our LUNA family cell counters in your comparison. It's worth the research!

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Specificity

Cell counting specificity

Even using multiple counters from the same model, the hardware used will be slightly different. Also, the size of the cell counting area matters too in how accurate the count will be. Imagine that you count a minimal area with few cells on it. Counting 9 or 8 cells results in a higher expected variability than a large area with either 345x10^6 or 300x^6 cells. The larger the area of counting and the sophistication of the algorithm used to correct hardware disparity, the less variability to expect.