Compensation flowjo12/3/2022 If your compensation matrix requires manual adjustments it can be a time-consuming process, and is often easier performed post acquisition on the analysis software as opposed to on the cytometer where time is billed. It’s possible that we have a very large panel or very little time for our experiment, and have not optimized the gate placement of our single stains for compensation, but rather quickly put something together to record, with the expectation that we will fine tune afterwards. There are times where it does make more sense to apply, or reapply compensation afterwards in analysis software. There may be extra steps that need to be done to accomplish this when a compensation matrix is created on analysis software. So, if you’re in a situation where you need to share FCS files with multiple people, be aware that the compensation matrix used may not be packaged appropriately with the FCS files you are sharing. This is not necessarily with a compensation matrix created in analysis software. Additionally, when compensations are set and applied on the cytometer, the FCS files exported should be packaged with that matrix and bring that matrix with them into analysis software like FlowJo. This gives us the ability to perform, on the spot, basic analysis of how our experiments performed. As trained here in the facility, we record our single stains, apply the compensation and then run our samples to get an accurate representation of our data, rather than running with no compensation applied. When performing a flow cytometry experiment on one of the analyzers, your first opportunity to perform compensation comes in the collection software, be this DIVA on the BD machines, CytExpert on the Cytoflex, or SpectroFlo as part of the unmixing, analogous to compensation, on the Aurora. Knowing the pros and cons of each of these features on both the cytometer and analysis side we can alter how we approach our experiments to get the best possible data. While we wouldn’t recommend users to do manual compensation, it is occasionally valuable to make manual adjustments to the matrix since compensation isn’t always perfect and some slight over or under compensation needs to be corrected. On top of this, there is also the ability to manually create the compensation matrix on both the cytometer side and in the analysis software. This can be confusing though, why have both, and when is it right to choose one over the other. In the process of performing flow cytometry experiments you’ll notice that while we almost always calculate the compensation matrix on the cytometer using the automated compensation tool, we also have the ability to do this in our analysis software such as FlowJo. #COMPENSATION FLOWJO REGISTRATION#Don't miss this opportunity to learn from the very best cytometrist of the UK ! Registration and more details are HERE.Ĭompensation in DIVA vs Compensation in FlowJo and a Look into the Future Places are limited to ensure maximum interaction. The course is supported by FlowJo and FCSExpress and delegates will have access to recorded tutorials as well as the data sets used in the course. They cover some of the common applications of flow - multicolour phenotyping (including compensation), cell proliferation (DNA cell cycyle and dye dilution), and funcional assays (apoptosis and calcium flux). This course is aimed at those new to cytometry or those who want to expand their capabilities. Is there one better than the other or do they complement each other ? Find out !įinally, the Royal Microscopical Society is running an interesting Flow Cytometry Data Analysis course from March 7th to March 10th. Additionnally, keep in mind that Kevin, Francisco and I are always available for sorts and trainings on either sites !įor this month FACS Tips, we are adressing the often asked question about compensation in DIVA vs Flowjo. As a consequence, the sorting schedule starting 2050 and + has been opened since he will undoubtedly take over Daddy Danny's Legacy.ĭanny is already back at the Agora with Mariela ! So the team is ready to help you in your projects. He is the proud and happy father of Adam Friedrich Labes, born on January 16th ❤️. I would like to announce that our beloved Danny is back at the Agora. FCF UNIL February 2022 (Compensation in DIVA vs Flowjo)Īs the snow ❄️ and the health restrictions □ are melting away, I hope you are feeling good in this month of February !
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