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Flow Cytometry

One For You, One For Me… How a Cell Sorter Works

Cell sorters do not operate by magic, even it looks that way. It’s about the application of physics, electronics, fast computers and formation of droplets. Whether you bring your cells to a flow core to be sorted or you sort them yourself, it important to know how the cell sorter works. Cells can be sorted…

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Crap in, crap out: Flushing Out The Problems in Your Flow Cytometry Data

“What Have You Done To My Cells??!!!” This cry of pain from researchers, frequently aimed at core facility operators, is heard after receiving incomprehensible data for an invaluable tube of cells. Equally baffling to the trained user of flow cytometric instrumentation is when data emerges that is either unreliable or inconsistent with the known properties…

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All for one and one for all – Fluorescence Minus One Controls

Ever done a multicolour flow cytometry experiment, run all your controls, done your compensation and then started to analyse your data and realised that you can’t work out where to put your gates? To err is human… When acquiring data on a cytometer, there can be measurement errors due to counting statistics, errors in processing…

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Time – The Forgotten Parameter in Flow Cytometry

In this webinar you will learn: why time is a forgotten but important parameter in flow cytometry how you can study a variety of temporal changes (from seconds to days in length) using flow cytometry. Summary: Time is one of the forgotten parameters in flow cytometry. It is highly used and has to be used…

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Cell Cycle Analysis by Flow Cytometry

In this webinar you will learn different methods for analysing the cell cycle by flow cytometry answers to common questions about using flow cytometry to study the cell cycle how to properly analyse cell cycle data. Summary: Analysis of the mammalian cell cycle was one of the first applications to which flow cytometry was put…

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There’s No Business Like Flow Business – Basics of Flow Cytometry

the basics of flow cytometry, including fluidics, optics and electronics. Summary: Flow Cytometry is a powerful technology that is quickly evolving.  It is important to understand how the cytometers work in order to get the most from your experiments.  This webinar will go through the basics of flow cytometry including fluidics, optics and electronics of…

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Take Control of Your World – Five Controls for Flow Cytometry

No, this is not a call for Geeks to take over the world – just a tiny part of it – the part that ensures success in all experiments or at least a good way to analyze them if they fail. There are all too many entry points for error and variability that can be…

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Intracellular Cytokine Staining: Letting It All Build Up Inside

Cytokines, those small proteins that modulate immune cell responses, once translated are normally secreted rapidly out of the cell. So, previously we could only check the levels of cytokines secreted in the supernatant, but we wouldn’t know which cell was producing which cytokine. But what if we had a way to keep the cytokines inside the cell?  Then we…

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Cell Proliferation Round 2 And Beyond: The Dye Dilution Method

Like the legendary fight between boxers Bowen and Burke in 1893, the cell cycle in some cells goes on and on..round 1, round 2, round 3, round 4…before the final bell is rung. Nucleotide analogs, like BrdU or EdU, are great for examining 1-2 cell cycle division(s).  In many studies though, for example the proliferation…

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Biosafety in Flow Cytometry – To Be or Not to Be…

Biosafety is one of those things many scientists don’t take seriously. I would guess, that like politics, there are 40% who believe biosafety is ‘over-emphasized’ and 40% who swear by biosafety. 20% are undecided. Needless to say, I’m on the side of biosafety. And here’s why: “CDC announced today that approximately 75 Atlanta-based staff are…

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Sorting Single Cells – What Do You Need to Consider?

Flow cytometer and cell sorter manufacturers have invested considerable resources to design instruments that are the “fastest in the ‘hood” either in terms of cells analyzed per second, or in total throughput. The general idea is the faster you can go, the quicker you can identify rare cells, and produce sorted populations containing large numbers…

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10 Useful Tips For Improving Your Sorting Experiments

After a very naïve start in flow cytometry thinking that published protocols will work without fault – needless to say, that did not work out in my favor – I realize now that following a few simple steps can go a long way, and will undoubtedly save you time in the end. So, here are…

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An Introduction to Gating in Flow Cytometry

What is one of the first things you do when you sit down at the flow cytometer and start looking at your cells? You start drawing polygons and setting gates. To the neophyte the gating process can look a little random – why do you exclude those dots but not these?  But gating in flow…

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Cell Proliferation Round 1: Using Thymidine Analogs With Flow Cytometry

Around and around the cell cycle goes, where it stops, nobody knows. Unless you have the right tools to analyze DNA content, that is. The DNA markers propidium iodide, Hoechst and DAPI are commonly used in flow cytometry to analyse a cell’s DNA content.  Although they are simple to use, they do have disadvantages. Figure…

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Seeing is Believing: An Introduction to Imaging Flow Cytometry

What if there was a way to take the power and speed of a flow cytometer and couple it with the resolution of a microscope? Imaging flow cytometry does just that! Flow cytometry is a very powerful tool for the complex characterization of cells and cell populations but flow cytometers can also be thought of…

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Getting in the Flow: Online Resources for Flow Cytometry

You are sitting in a seminar when you glimpse it:  the figure that beats all other figures –  a beautiful contour plot – and you realise that a similar figure is what you need to publish your paper in a high-ranking journal.  You know the basics of flow cytometry, but admit you need a little…

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Herzenberg and the Invention of the FACS Machine

The flow cytometer that we have all grown to know and love may have only come into its own in the 1990’s, but who would have known that the first cell sorter was invented as early as the 1950’s? With the recent death of one of the key developers of fluorescence activated cell sorting (FACS),…

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“Pick ‘n’ Mix? A Basic Guide to Commercial Flow Cytometers”

  So having read our article on how a cytometer works, surely the next question is ‘what’s the right flow cytometer for me?!’ Basic Components of Flow Cytometers We know that at their most basic level, cytometers are made up of 3 main components: A fluidics system to transport and focus the cells past an…

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Evolution Flow: The Historical Background of Flow Cytometry

For those of you who print out the results that you have acquired from the Flow Cytometer to stick in your lab book, did you know that inkjet printers and cytometers have a shared history? Flow Cytometry is less than 50 years old and machines today still use some of the same principles as the…

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