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3 Action Steps You Can Take Right Now To Improve Your Flow Cytometry Reproducibility

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Reproducibility is a state of mind. It’s not one simple thing that you do that will make all your data more reproducible, it a shift in the way one thinks about and perform experiments. With the emphasis on rigor and reproducibility in science, it’s very important that researchers start putting into place everything they can do to help improve the quality and reproducibility of there data. Learn 3 action steps that can be taken to enhance experimental reproducibility.

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3 Questions You Should Be Asking About Flow Cytometry Controls For Your Experiments

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Controls are an incredibly important part of your flow cytometry experiments. If not done correctly, poor controls will waste time and money. But with proper care, high-quality controls will result in high-quality data. Just be sure to ask yourself these key questions, should you be using isotype controls, do you have a quality control procedure in place, and are you following the 3 cardinal rules of compensation.

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3 Ways To Improve Flow Cytometry Troubleshooting

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A lot of the troubleshooting is focused on fluidics issues. If you sit down and think about your workflow, and how you might want to add a couple of little tweaks here and there which will ultimately help you improve the quality of your data as well as aid you in identifying issues before they become problems your troubleshooting will be much smoother. Consider these three things, what do you before you start collecting data, ensure you have appropriate plots of time vs fluorescence for each of the lasers your using and apply appropriate gating procedures.

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Using Begley’s Rules To Improve Reproducibility In Flow Cytometry

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Cell sorting is a combination of a numbers game (Recovery), quality of output (Purity) and speed. For any experiment, the end goal is going to be measured by these three characteristics, and as soon as one of these measures is more heavily favored, the other two must be compromised in some manner.
When designing a sorting experiment, start with the question of what will the cells be used for after sorting, and how many cells will you need for those experiments? That will set the minimum recovery that is needed. The second question is how pure do you need the cells? The requirements of the downstream assay will also dictate the purity needed.

The cell type being used will, in part, dictate the speed of sorting. Smaller cells can be sorted faster because a smaller nozzle can be used.

When you start a cell sort it’s important that you are aware of the downstream analysis and assays that you want to run. This will determine how you perform the sort and how you determine if your sort was successful or not.
Successful cell sorting involves balancing recovery, yield and speed. What do these three terms mean and what influences each of these factors?

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