Flow Cytometry
The History and Future of Fluorescent Labels: We’ve Come a Long Way, Baby!
If you’ve been keeping up with our recent series of articles, welcome back! If not, you can catch up on how fluorescence works or what not to do with your flow experiment. In short, we have been discussing fluorescent labels and their role in flow cytometry. Today, I’ll round out our discussion by touching on…
Read More 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.…
Read More Lighting the Way: Understanding Flow Cytometry Fluorophores
As science is becoming more interdisciplinary, the tools we use to answer questions are also crossing party lines. Case in point: flow cytometry. Once a tool only used by “real” immunologists, flow cytometry is fast becoming a method by which numerous questions can be answered, from the length of a cell’s telomeres, to the state…
Read More Analyzing Cell Signaling with Flow Cytometry: Go with the Flow
Phosphorylation Equals Cell Signaling! How do cells communicate and respond to their environmental cues? This question has been on the hot list for scientists ever since the discovery of the cell. Cells use signaling cascades based on biochemical reactions to deliver or receive messages. How cool is that? The major secret of cell signaling was…
Read More Detection of Apoptosis by Flow Cytometry: To Be or Not to Be
Sometimes only a small subset of a cell population will show apoptotic features making flow cytometry an excellent way to identify and quantify them. A previous Bitesize Bio article showed how flow cytometry can detect apoptotic hallmarks. More than 30 different dyes can be used to detect apoptosis. It is also true to say that…
Read More Incorporating New Dyes to Simplify Panel Design: Accelerating Reagent Availability
In this webinar you will learn how custom reagents can aid you in flow cytometry panel design—without long waits for generating these components. We will cover: How quality custom reagents in small sizes will make your panel design easier How you can receive these reagents fast and without traditional wait times for manufacturing How you…
Read More 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…
Read More How to Destroy your Flow Cytometry Data in 3 Easy Steps: Snap, Crackle, and Pop
While many scientists are methodical and precise, some of us like to live on the edge. Read a protocol all the way through? No thanks, I’ll take my chances and guess what concentration of HCl I should use. Label my tubes with the correct content? Puh-lease – it’s much more exciting deducing which is which…
Read More 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…
Read More 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…
Read More Ensure Reproducibility: Control for Lot-to-Lot Variation of Antibodies
When starting a long-term experiment, you need to take a lot of things into consideration (availability of cells, reagents, planning time points), but do you ever think about your antibodies? If you buy an antibody from a manufacturer, run out half way through the study, and buy the same antibody again, have you thought about…
Read More 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…
Read More 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…
Read More A Simple Way to Measure T cell Killing Activity In Vivo
Have you ever wondered what a cell’s life is like? Do they constantly communicate to each other or do they just go on their own daily business? There is an easier way and it involves super advanced molecular “crayon” technology.
Read More 2015 Staff Picks: Flow Cytometry
There’s a joke in here about our editorial team going with the flow, but I can’t quite seem to make it work (feel free to leave suggestions in the comments). Despite my inability to make a joke count, I do hope that you find these articles amusing and useful. Jen Redig, Managing Editor Spot the…
Read More Cell Cycle Analysis by Flow Cytometry: Flowing your Way through Life’s Cycle
Over the past few decades the mammalian cell cycle has been well documented. Although there are lots of checkpoints as cells move through the cycle, we can very simply divide the cell cycle into three stages according to the DNA content in the nucleus. When cells are either quiescent or not dividing they have the…
Read More Gateway to the Cellular Kingdom: Cell Fixation and Permeabilization Techniques
One of the much sought after question asked by many researchers worldwide is – “What is the gene expression profile of a single cell within a heterogenous pool of cells?” While mass cytometry is the current ‘hot’ methodology for single cell analysis, the good old flow cytometry can help us perform rapid analysis of single…
Read More 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…
Read More Locating Your Cellular Apoptosis Squad: Annexin V Staining Assays
In real life, cells are instructed to commit suicide for the greater good of the organism. The programmed cell death (apoptosis) is important during development of a multi-cellular organism. A good example you will appreciate is the dis-appreance of the tail from a tadpole as it turns into a frog. On the reverse, the lack…
Read More 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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