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6 Tips For Applying The Right Statistical Test To Your Flow Cytometry Data

6 Tips For Applying The Right Statistical Test To Your Flow Cytometry Data

Flow cytometry data are numbers rich. Data from experiments can be population measurements (percent of CD4+ cells, for example), or it can be expression level (median fluorescent expression of CD69 on activated T cells). Many times, researchers are content to show histograms to illustrate their point after a flow experiment. This approach misses the opportunity to take that content rich data and extend the analysis into a statistical analysis. To properly perform statistical analysis, the first step is to understand the hypothesis. The hypothesis will guide the statistical analysis, identifying the correct test to be performed. There are several things that need to ...

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7 Tips For Measuring And Reporting Apoptosis By Flow Cytometry

Cell death is a fact of biological life.  How, when, where and most importantly, why cells die, can have huge biological consequences on the path an organism may take. Apoptosis, or programed cell death, can result in a selective advantage for an organism. Fingers, for example, are the result of apoptosis of cells during development. Next to immunophenotyping, measuring apoptosis using flow cytometry is one of the most common assays. It may be because of the many different ways to measure the process, many of which can be easily performed in a high-throughput manner, or combined with other assays to determine if specific cellular ...

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If You Don't Know This About GFP, FITC, And PE, You Might Publish False Flow Cytometry Data

When we learn about fluorescence, the first thing we are told is that fluorophores emit photons that are higher wavelength than the photons that they absorb. What this specifically refers to is the stokes shift, which results from non-radiative energy transfer during the fluorescence process. When a photon is absorbed by a fluorophore molecule, some of the resultant energy is lost in molecular vibration and movement (among other things) so that the energy released after fluorescence is lower than the energy absorbed. Since wavelength is inversely proportional to energy, this lower output energy light is higher in wavelength than the input ...

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How To Do Phospho-Flow Cytometry

I often have researchers come into the core wanting to look at the activation and downstream signaling events that occur in different immune cells. These events occur in response to signals such as cytokines, chemokines, various receptor ligands, and the engagement of the T cell or B cell receptors. The signaling events are also characterized by the initiation of several phosphorylation events. Measuring Phosphorylation Events When this is the case, I recommend that the researchers set up a phospho-specific flow cytometry, or phospho-flow, experiment. These types of experiments measure the phosphorylation state of intracellular proteins at the single cell level. Phospho-flow allows for the ...

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5 Mistakes Scientists Make When Doing Flow Cytometry Proliferation Experiments

5 Mistakes Scientists Make When Doing Flow Cytometry Proliferation Experiments

Measuring cell proliferation can be done in a number of ways. There is the below tried and true method of counting cells. This straightforward assay can help determine if the cells are proliferating and by comparing counts. Here, a researcher can determine that the experimental treatment is increasing cell growth. A second method of measuring proliferation involves using a radioactive tracer like 3H-Thymidine. In this assay, the amount of the isotope taken up by the cells correlates to the amount of DNA synthesis, and therefore growth. Of course this requires using radioactivity and all that entails. A third method is using a something ...

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Testimonials
Robert Fultz Baylor College of Medicine
I have much more confidence in the quality of data that I’m able to get from my samples after taking the Expert Cytometry, Complex Data Analysis course that was offered in Houston this year. I found the curriculum, the way it was taught, and the hands-on learning to be hugely helpful. I am very new to the world of flow cytometry so proficiency in even the most basic data preparation and analysis is critical for me and this class really put ahead of the curve. Additionally, using the techniques I learned in the course, I re-evaluated some data that I previously collected and found a critical error with the way the samples were run. The error would not have been detected without the knowledge I acquired in the course. I strongly recommend this course for anyone doing even simple FC analysis.

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I attended an Excyte course at the University of Iowa. The course was extremely detail-oriented, allowing attendees to work through the process of choosing the correct flow cytometry flurophores and reagents for their particular experiments. I learned a lot from the discussions on flow cytometry controls, which helped me figure out which ones to use in my own research.  I found the instructor very approachable and willing to stay after the lecture for a long time to answer questions and give additional resources.

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I invited ExCyte to Lilly to teach a two-day comprehensive course; ranging from the fundamentals of flow to the experimental design of assays specific to what I think a lot of biotech companies are doing these days. Afterwards, several of my users wondered why I didn’t bring in a course like this earlier! Our instructor, Tim Bushnell, Ph.D., educated my users with the background they needed to design their assays most effectively. I immediately noticed the positive impact he had when my users began to ask more informed questions during the planning stages of their experiments. As a busy core manager, it’s difficult to find the time to prepare such a well designed course like ExCyte offers while trying to stay focused on innovation and maintaining a competitive advantage in the fast paced industry environment. I’m grateful to the ExCyte team for helping me to keep things moving forward at ...

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Excyte provided a wonderful course in Boston.  Feedback from attendees has been very positive and individuals want more. I believe there is great benefit to advanced courses in Flow Cytometry as number of fluorescence antibodies and analysis parameters keeps increasing. Between the multi-institutional CyTOF, and my labs 32 parameter Astrios, we are moving into some very high-end and cutting edge science. The expertise that the ExCyte team brings to these courses is incredible. It is amazing how some individuals such as ExCyte can take very complex materials and make them appear simple. In addition, I am and always will be a strong advocate of learning by doing. ExCyte's course structure allows the student to be involved and interactive. This allows for greater ease of translation of the learned material into the student's experiments. Great courses from great people.

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As a core manager, ExCyte has been great to work with.  It's often difficult to ensure clients are getting the information and education they need to perform their experiments effectively.  We have worked with Carol and Tim several times to deliver effective educational programs to our users.  They have a broad offering of classes and are willing to work with us to tailor the topics to our needs.  Client feedback has been very good which is a testament to the knowledge and expertise of the people behind ExCyte. I also like their new series of webinars which are inexpensive, informative and allow users to gain valuable knowledge one hour at a time.

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