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Why Flow Cytometry Fluorescence Compensation Is Critical For Quality Measurements

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Fluorescence compensation is one of the more difficult, understandably confusing, and misunderstood aspects of flow cytometry. Understanding what compensation is, why it is necessary, and what to expect when using it, are critical for generating useful and high-quality data from flow cytometry experiments. The definition and mechanics of flow cytometry compensation and the critical concept that compensation’s most basic principle relies on are discussed here.

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12 Flow Cytometry Terms And Definitions Most Scientists Get Wrong

Flow Cytometry Terms And Definitions | Expert Cytometry | flow cytometry meanings
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The most important part of executing a flow cytometry experiment correctly is actually understanding what you are doing. This means you must understand the terms and definitions that are critical to the field of flow cytometry. You must also be able to communicate your methodologies and results intelligently. To this end, we have compiled this list of the top 12 most commonly unknown or commonly misunderstood flow cytometry terms and definitions.

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Why You Need To Use FMO Controls For All Multicolor Flow Cytometry Experiments

multicolor flow cytometry training | Expert Cytometry | fmo control flow cytometry
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FMO controls are samples that contain all the antibodies you are testing in your experimental samples, minus one of them. When analyzing the minus, or left out parameter in an FMO control, you give yourself a strong negative control to work with. It’s a strong negative control because the left out marker in the FMO control allows you to take into account how the other stains in your panel affect the respective minus parameter. Many flow cytometry gates are difficult to define. This is especially true when you’re looking at activation markers within a continuum or accounting for the large data spread that occurs when compensating a 10+ color experiment. The only way to convince reviewers that your gate is in the proper place is by using FMO controls. Here’s why you need to use FMO controls for any multicolor flow cytometry experiment and how to prepare these controls properly.

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How Droplets Are Charged And Drop Delays Are Determined During An Electrostatic Cell Sorting Experiment

cell sorting techniques | Expert Cytometry | cell separation techniques
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Electrostatic cell sorting is a complicated process that continues to be improved. It can be a struggle to understand exactly how all of the sorting components coalesce to accomplish the cell sorter’s tasks. For many scientists, the most difficult parts of the sorting equation are how droplets are charged and how drop delays are calculated. By understanding these two things, you will be in a better position to set up a successful fcell sorting experiment, which will help you achieve high sort recovery values, allowing for the accurate analysis of your cells and more cells to work with for your downstream experiments. Here’s how cell sorting droplets are charged and how cell sorting drop delays are determined.

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What Is The International Cytometry Certification Exam (ICCE) And How To Pass It

ICCE and Cytometry Certification | Expert Cytometry | clinical cytometry society
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The International Cytometry Certification Exam was developed over a period of several years. The goal was to ensure a base level of flow cytometry knowledge in certificate holders. Some cytometrists have deemed the ICCE as unnecessary. Others have voiced concerns about the specialization of certain exam subsections. However, despite these concerns, the ICCE is here to stay. The exam and certification process as a whole has the support of multiple companies that are providing training, as well as the support of ISAC, ICCS, and the Wallace H. Coulter Foundation. In the end, the most telling test of the value of the ICCE will be when flow cyotmetry job postings in basic research start asking for the certification. This day may not be too far away.

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