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For those new to flow cytometry, compensation is confusing at best and terrifying at worst. Likewise, those who have been doing flow cytometry since the analog ages may be holding on to practices that, while suited to the analog instruments, should be left to the annals of history. As such, a lot of time is spent discussing compensation and the best practices for this critical process. There are 3 rules that guide proper compensation, and they’ve been written about extensively since they first appeared in the “Daily Dongle” in 2011. Here, we will review the classic rules and expand upon the tacit assumptions required to fulfill them.

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Congratulations, your grant has been funded! Next comes generating data and publishing papers. What was that hypothesis again? It must be in the grant somewhere, right? To avoid even the appearance of HARKing — Hypothesizing After The Results Are Known — it is important to start the statistical analysis process even before the first experiments are performed. This process consists of 5 steps: setting the null hypothesis, establishing a threshold, performing the experiments, performing the statistical tests, and communicating the results. Walk with us as we discuss these steps in an example workflow.

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