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Flow Cytometry

Cell Cycle Analysis by Flow: DNA Stains and Beyond

While you can observe mitotic cell cycle progression using immunofluorescence, flow cytometry is a great tool to delineate details that aren’t apparent by chromosomal morphology alone. DNA stains are a great way to get a general idea of what your cells are up to. There are also a number of other stains you can use…

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A Biologist’s Guide to Choosing Your Fluorophore Palette

You may notice that nature is full of vibrant, even fluorescent, colors. The human eye detects wavelengths ranging from 390-700 nm and our perception of colors is actually a narrow part of the light spectrum. Other organisms can detect color from a wider spectrum. Why do colors exist? Arguably, colors are communicative, from tropical fish…

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Flow Cytometric Apoptosis Assays for Cell Death

Apoptosis, often called programmed cell death, is a carefully regulated process that is part of normal development and homeostasis. Apoptosis is morphologically and biochemically distinct from necrosis, which is conversely called accidental cell death. Dysregulation of apoptosis is implicated in disease states such as cancer, autoimmune disease and degenerative conditions. Apoptosis consists of an orderly…

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Top 5 Tricks for Using FlowJo

Are you planning to do cellular immunology research?  Then chances are you will be introduced to the flow cytometer –  “a modern immunologist’s best friend.” This modern magic box is a highly versatile machine packed with cutting-edge fluidics and photonics (lasers). Combined with the monoclonal antibodies conjugated to fluorochromes capable of emitting light signals from a…

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Introducing CyTOF: Cytometry of the Masses

Flow cytometry remains unparalleled as a single-cell analysis technology.  The ability to analyze 14 or more fluorescent parameters on a million cells or more allows for detailed understanding of complex biological processes. The Problem With Traditional Flow Cytometry One limitation of flow cytometry is the reliance on fluorescent tags.   Even with careful panel design, loss…

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Mass(ively powerful) Cytometry

Mass Cytometry is a relatively new technology which has recently featured in many high-impact journals. You may have read about instruments including the CyTOF, CyTOF2, and more recently, the Helios. With these instruments becoming more widespread, you might find yourself asking, what is mass cytometry, and what can it do for you? The Basis: Conventional…

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Ain’t no mountain high enough: Summit Software

The town of Fort Collins is situated in the foothills of the Rocky Mountains in Colorado and it is those mountains that give inspiration to the name of the Beckman Coulter acquisition program Summit. Summit software started out as the software to run the MoFlo (Modular Flow) high-speed cell sorter that was designed by Ger…

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Remote Cytometry: Help from beyond!

The idea of accessing one computer from another is long established. Unfortunately, we often have visions of hackers sneaking in and stealing our data when we have most to lose. However, this type of technology can aid us in a lot of applications and to those of us who work in cytometry the benefits are (somewhat) clear. No More ‘Fail’ Moments Many researchers know the dread of…

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How a Flow Cytometer Works: A Look Inside the Magic Box

A flow cytometer is a device used to illuminate objects and capture and quantitate light emitting from these objects.  The “objects” are normally single cells dispersed in a medium, but could very well be polystyrene beads, cell fragments or debris, or even large molecules. So, What’s in the Box? Using your highly tuned powers of deduction, you…

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Sorting Large Cells and Materials by Flow Cytometry

Flow cytometers and cell sorters were designed with blood cells in mind. This means that commercial cell sorters are optimized for sorting cells typically smaller than about 20 µm in diameter. However, it turns out that many cell types, including those of mammals, are larger than 20 µm. So what are your options if you…

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The Exciting (and Emitting) World of Fluorescence

Flow cytometry is a fluorescence-based technology, as is fluorescence microscopy and confocal microscopy. Fluorescence is fundamental to how a cytometer gathers data, but I am often surprised, as a core manager, at how little new users know about the process of fluorescence. So, this is where I always start the training process. Let’s get physical…

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Catching Greatness: Measuring Cellular Degranulation

One of the key characteristics of cytotoxic cells (i.e. CD8+ T cells, natural killer cells) is the presence of pre-formed cytoplasmic lysosomal granules. These structures house perforin and granzyme; two molecules that are essential for the lysis of target cells. Upon effector cell activation, granules are polarized toward the target cell and the contents are…

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An Introduction to Biomarkers

During the past decade or two, the field of biomarkers has witnessed immense development. The complexity of the field mirrors its multi-faceted applications, and it is easy to get confused with the different terms used in different contexts. Hopefully, this article will help make things clearer! What is a Biomarker? The word ‘biomarker,’ has more…

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