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The Difference Between Purity, Single Cell, And Recovery Cell Sorting Techniques

Written by Michael Kissner If you have experience sorting different kinds of cell types on a droplet sorter, you may have noticed that sorting efficiency often seems closely tied to the cell and sample type. For instance, you may have experienced a better outcome, such as a higher efficiency and more cells recovered, after sorting lymphocytes versus sorting an adherent cell line. There are some fundamental concepts that underpin this phenomenon, and understanding them will help you perform better cell sorting experiments. What Is Flow Cytometry Cell Sorting Efficiency? Sorting efficiency, in fundamental terms, is a real-time measurement, generated by the instrument, of how ...

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12 Flow Cytometry Terms And Definitions Most Scientists Get Wrong

Written by Tim Bushnell, Ph.D. 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. As a scientist, you should not just place your faith in a specialized technician. You should not blindly agree with the data you see in front of you without you yourself knowing what ‘logicle scaling’ means, what ‘differential pressure’ means, what happens when you change the differential pressure on an instrument, and so on. You must also be able to communicate your ...

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How To Compensate A 4-Color Flow Cytometry Experiment Correctly

Written by Tim Bushnell, Ph.D. Compensation in flow cytometry is a critical step to ensure accurate interpretation of data. It is also one of the areas that’s steeped in mystery, myths and misinformation. Before jumping into the best practices for compensation of flow cytometry experiments, it’s good to show what NOT to do when performing compensation. Manually adjusting the compensation values based on how the populations look, or so-called ‘Cowboy Compensation’ (thanks to Joel Sederstrom for the term), is not the correct way to determine proper compensation. For example, review the following figure, and ask yourself what is the best compensation value? This figure ...

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3 Flow Cytometry Gates That Will Improve The Accuracy Of Your FACS Data Analysis

Written by Tim Bushnell, Ph.D. Flow cytometry data analysis typically involves a step where a series of gates are defined to identify the population of interest. The development of robust gating strategies, and the communication of these strategies can be complex. Figures in papers often only tell part of the story. In the world of multicenter studies and the need for improved reproducibility, understanding the specifics of each gating strategy is critical. As Maecker et al., pointed out in this article, the coefficient of variation due to data analysis went from 20.5% when performed at individual labs, to 4% when performed centrally. With ...

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What Is Flow Cytometry Light Scatter And How Cell Size And Particle Size Affects It

Written by Mike Kissner Light scatter is unfortunately one of the more misunderstood concepts in cytometry. When you first learn flow cytometry, you were likely told the misleading phrase, “Forward scatter signal intensity is proportional to cell size, and side scatter signal intensity is proportional to cell granularity.” The short story here is that, yes, this can be the case when we’re dealing with typical cells (consider blood granulocytes, which are bigger than lymphocytes, have more intense FSC signals; granulocytes are more “granular” than lymphocytes and thus have more intense SSC signals). However, the truth is that light scatter complexity is belied by ...

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WELCOME TO EXCYTE

ExCyte is the world’s leading flow cytometry training company. Whether you are a core manger looking to keep your users on the cutting edge, or a business professional looking to enter into this rapidly growing market, ExCyte can help. We have the information and experience you need.

ExCyte is currently booking beginner and advanced flow cytometry courses around the world. Click on the Calendar box above to see where we will be next.

Testimonials
MedImmune purchased a site license of Expert Cytometry’s Mastery Class. We have over 100 scientists using the program and the feedback has been phenomenal. I’m amazed at how much the scientists are coming to me with questions that are much more advanced than what I used to get before having access to the Mastery Class. I’m now able to focus my attention, and that of my staff’s, to more advanced questions and inquiries specific to the researcher. We are also having less trouble with the mistakes that new users make and spending less time dealing with troubleshooting basic issues. As the core director at MedImmune, I definitely recommend the Expert Cytometry Mastery Class to all cores, particularly in a corporate setting, where we simply do not have the time to train all of the new users in best practices. My core is running 10 hours a week, ...

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Johnna Wesley, Ph.D.Senior Manager, Novo Nordisk
Expert Cytometry provided an excellent course to our group. The course was over two days. On day one, the instructors provided a great overview of core concepts of flow. On day two, we were able to customize the course to our needs. It was very valuable and I would recommend that if your group is starting to use flow more heavily, that you bring Expert Cytometry in for a course. There are nuances that I never knew and it immediately helped our group perform better experiments.   

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I was (and still am for the most part) completely ignorant of the details of flow cytometry. The Expert Cytometry team provided explanations and helped me understand the technology to the point where I was able to intelligently and efficiently manage both of our contract flow labs. Expert Cytometry provided independent assessments of the quality of the work and data from the labs that allowed me to make good decisions regarding the results. Finally, I was always amazed at how quickly Expert Cytometry responded and completed the analyses when we were on short timelines. We continue to work with ExCyte and will for the foreseeable future.

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Our group at BlueBirdBio was relatively new to flow. We brought ExCyte in for a basic course, and also to help train us on best practices for our new flow cytometer. The level of flow cytometry knowledge ExCyte brought with them was amazing. I can’t believe how much experience ExCyte has and how much they bring to the courses they provide. ExCyte was able to answer all questions with real world examples. The course brought the theory of flow to the real world and helped us understand the concepts more easily. ExCyte was professional, courteous, and it almost felt like they became part of our group while we were working together. I know I can ask ExCyte any questions whenever I need them and I hope to continue to work with them as we move forward. I have no doubt we’ll bring ExCyte back for another course in ...

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We worked with Expert Cytometry for one year, utilizing them for consulting work and writing technical documents for several of our flow cytometry products. Working with Tim Bushnell at Expert Cytometry was great. Tim is efficient and highly technical. His 15+ years of experience really shows, as he brings a level of detail to the work that we had never experienced from other consultants. There are a lot of so-called flow cytometry “experts” out there, but having worked with numerous ones, we have found the group at Expert Cytometry to be the most knowledgeable. As we continue to expand our flow cytometry portfolio, I will definitely work with Tim and his group at Expert Cytometry again.

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