The Daily Dongle

Three Rules for Compensation Controls

First and foremost, there must be a single stained control for every parameter in the experiment! In addition, there are three rules for “good” compensation controls: 1) Controls need to be at least as bright or brighter than any sample the compensation will be applied to 2) Background fluorescence should... Read more →

Transformation Differences

One of the most frequently questions I'm asked is, "Why does the data look different in FlowJo as compared to DIVA?" It is a result of the biexponential transformation. DIVA calculates the "best" transform and typically uses a width basis around -100 by default. Therefore, the first two decades are... Read more →

FMO vs. Isotype Controls

In addition to compensation controls, there are several other controls that can, and in most cases, should be used to help resolve issue in staining. Fluorescence Minus One (FMO) controls can help identify gating boundaries, isotype controls can help identify staining issues and unstained controls show you the background or... Read more →