Techniques / Flow Cytometry

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Chromosome Analysis by Flow Cytometry

In most people’s minds a flow cytometer can sort, view and count cells e.g. lymphocytes, thymocytes, cultured cells and even non-mammalian cells such as yeast or bacteria. However, in reality, a flow cytometer is capable of providing information about any particle as long as it has detectable fluorescence. This fluorescence may occur either inherently or […]

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In Flow Cytometry 27th of April, 2017
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How to Store Your Reagents, so They ‘Do Exactly What It Says on the Tin’

Your reagents should do ‘Exactly what they say on the tin.’  This only happens though if you look after them in the way the manufacturer states on their data sheets. We have all been guilty of using reagents past their expiration date.  Usually we can get away with it, but there are a few things […]

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In Flow Cytometry 11th of April, 2017
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Are Quantum Dots Any Good for Flow Cytometry?

What Are Quantum Dots? Quantum dots were discovered in the early 1980s. However, it was not until the late 1990s that their use in biological applications was suggested.1 Quantum dots are semiconducting nanocrystals made of artificial atom clusters. Their size generally ranges from 2 to 20 nm. Size is crucial for their physical properties because […]

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In Flow Cytometry 23rd of March, 2017
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How to perform cell synchronization in specific cell cycle phases

The cell cycle has been very well documented over the years because of its dysregulation in diseases such as cancer. Many different processes contribute to cell growth and replication, which is ultimately controlled by a series of tightly controlled cell cycle phases. For some areas of research, especially within drug discovery and cancer research, cell synchronization in […]

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Hierarchical or Boolean Gating: Which One to Choose?

A flow cytometer collects the events you are interested in, and also ‘sees’ every event that goes through. This includes debris and even bits in your buffers. As cytometrists, we gate our cells to exclude unwanted bits and to focus on the sub-populations that we are interested in studying. There are two main ways of gating […]

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In Flow Cytometry 14th of February, 2017
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How Fluorescent Molecules Work: Shine Bright like a Diamond

Fluorescence is one of the most important and useful tools in a biologist’s toolbox. In biology, nearly every field, from physiology to immunology, uses fluorescent molecules (aka fluorophores) to detect proteins. However, the specific science behind how fluorescence works can be confusing or overlooked. Have no fear! In this article, we break down key points of […]

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In Flow Cytometry 27th of January, 2017
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Hydrodynamic Focusing in Flow Cytometry

If you have sorted samples or phenotyped cells by surface expression of proteins, you’ve probably wondered how each cell is sorted or phenotyped in a flow cytometer? This question seems trivial, but in reality it took a while for engineers to figure it out. Before I get into today’s topic on “hydrodynamic focusing,” I’ll walk […]

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In Flow Cytometry 10th of January, 2017
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Multiplex Cytometric Bead Array: The ABCs of CBAs

Multi-parameter data acquisition is key to the modern era of science research. I, for one, wish every single experiment that I design would give me the maximum amount of information. For example, in cell biology and immunology, we want to capture as much information (be it cytokines/hormones/chemokines) as possible about a given cell population. Of […]

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In Flow Cytometry 13th of December, 2016
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Demystifying the Flow Cytometry Optics System: A Peek Under the Hood

To many users, the flow cytometer is a magic box: put in cells, get out data. You click the button to tell it which colors to look at without much thought about how the machine does this. However, not all fluorophores are created equal—some configurations might exclude the spectrum you’re really looking for. Here’s a […]

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In Flow Cytometry 24th of November, 2016
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Corralling Your Cells: How to Gate in Flow Cytometry

Flow cytometry. Some people love it—most hate it—but all can agree that it is one of the most powerful analytical tools immunologists possess. Here’s a quick refresher: as the name suggests, flow cytometry measures the physical and chemical characteristics of cells. This is accomplished by fluorescently labeling cell surface markers/proteins using antibodies conjugated to fluorophores. […]

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In Flow Cytometry 27th of October, 2016