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Welcome to the blog
The content posted here is intended to help and is meant to be as accurate as possible. There will inevitably be mistakes or gaps in the knowledge. This is not peer-reviewed science, but is evidence-based as much as possible. If you see a factual inaccuracy or disagree, add a comment or send in a correction. Also, while some of this content tries to push the boundaries of what we know about how to do flow, it is not intended to claim invention credit. If you feel a reference has been overlooked, please let me know.
AutoSpill 2: Spectral Boogaloo
AutoSpectral is a new way of getting better unmixing.
Jun 301 min read
Stabilizing your unmixing: drift
Today's post is meant to illustrate why I do not recommend running new samples using the same unmixing matrix across multiple days.
May 122 min read
Stabilizing your unmixing
Today's post pulls from a lot of previous posts on this blog to show how to effectively eliminate batch effects in your flow from an experimental side. Stabilizing the unmixing is a big part of this.
Apr 285 min read
The peculiarities of Phenograph
Phenograph is one of the more popular clustering algorithms for flow cytometry.
Apr 145 min read
Unmixing: the tortoise beats the hare
Today's tip: when running your single-color controls, take it slow.
Mar 314 min read
Creating and using settings on the Aurora
Create settings to ensure reproducible results across experiments.
Mar 179 min read
Stabilizing master mixes: what doesn't work
If you don't put in the legwork of testing the mixes for your panel, expect to run into problems.
Mar 32 min read
Stabilizing master mixes
How to prepare and store antibody master mixes stably.
Feb 178 min read
Simple tips for reducing errors with panel prep
Running large panels consistently is all about error prevention.
Feb 36 min read
“Rescuing” GFP
Why do we lose GFP and other fluorescent proteins when we fix and permeabilize cells? How can we stop this?
Jan 206 min read
Overnight staining: tetramers
Does overnight staining help for antigen-specific TCR detection with tetramers? Not so much.
Jan 63 min read
Fluorophores for nuclear staining
What's the best type of fluorophore to use for targets that are in the nucleus of the cell?
Dec 16, 20243 min read
Dealing with multiple autofluorescences
Our samples, more often than not, contain autofluorescence (AF) that varies not only in intensity but also in aspect.
Dec 2, 20248 min read
How to: Run plates on the Cytek Aurora
Today I'm going to share my tips for successfully running lots of samples on the Aurora without having to babysit the machine.
Nov 25, 20245 min read
Five ways to extract autofluorescence (and store it)
Let’s start by looking at the autofluorescence extraction methods available to us, their advantages and drawbacks.
Nov 18, 202412 min read
Unmixing: should you use a universal negative?
When unmixing, we have the option to use either an internal negative population or a completely independent "Universal Negative".
Nov 11, 20244 min read
Autofluorescence in conventional flow cytometry
Let's take a step back and look at how autofluorescence is handled in conventional flow.
Nov 4, 20247 min read
ClusterID update
Thanks to franpcozar for highlighting an issue with ClusterID . The script was failing when the input clusters were named with...
Oct 31, 20241 min read
Backgating: What is it and why will it help?
Backgating is a simple, useful technique in flow cytometry that can help you figure out where your cells are, and thus optimize your gating
Oct 28, 20243 min read
Autofluorescence: what not to do
Today let’s look at something I think you shouldn’t be doing with AF extraction in spectral flow cytometry.
Oct 21, 20244 min read
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