Colibri Cytometry

Today's tip: when running your single-color controls, take it slow.

Create settings to ensure reproducible results across experiments.

If you don't put in the legwork of testing the mixes for your panel, expect to run into problems.

How to prepare and store antibody master mixes stably.

Running large panels consistently is all about error prevention.

Why do we lose GFP and other fluorescent proteins when we fix and permeabilize cells? How can we stop this?

What's the best type of fluorophore to use for targets that are in the nucleus of the cell?

When unmixing, we have the option to use either an internal negative population or a completely independent "Universal Negative".

Backgating is a simple, useful technique in flow cytometry that can help you figure out where your cells are, and thus optimize your gating

You've designed your big flow panel--a great accomplishment! But wait, there's more work to do before you use it.

What's the best way to stain for human CCR7?

What are the benefits and drawbacks of staining at higher temperatures?

Cell-based controls are inherently multi-colour controls. Understanding this is key to getting good unmixing.

Why do we want multiple controls? The most common reason, I expect, is that you'll have a control that you aren't sure will work.

CellBlox: a blocking solution for working with NovaFluors

The issue: Interactions between Brilliant dyes on antibody conjugates.

Monocyte Block destroys your cells. When used intracellularly, Monocyte Block reduces transcription factor staining.

Let's discuss Fc block, which is what most people think of when we talk about "blocking" for flow cytometry.

Today we're looking at the most basic of blocking reagents: generic protein.

In this series, I’m going to review blocking reagents and how to use them for best effect.