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Strengths And Weaknesses Of Isotype Controls In Flow Cytometry

Written by Tim Bushnell, Ph.D Controls are critical for minimizing the effects of the variables in a scientific experiment so that the effect of the independent variable can be accurately measured. In flow cytometry, there are a host of important controls necessary to properly interpret data generated in these experiments. Some of these controls include compensation, fluorescence minus one (FMO), stimulated and unstimulated, reference, and controls. When it becomes time to publish, the proper use of these controls is critical in convincing the reviewer and reader that the data has been properly analyzed. The isotype control is an experimental control where a sample ...

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How Flow Cytometry Converts Photons To Digital Data

Written by Tim Bushnell, Ph.D How exactly does a photon of light become an electron, and eventually a number in the FCS listmode file? In flow cytometry, cells are labeled with fluorescent tags, passed by an excitation light source, and the resulting emitted photons are collected to become the data that identifies characteristics about the cells in question. These characteristics can include what proteins are expressed, the cell cycle state, the phosphorylation state of a protein, the calcium state of the cells, levels of RNA expression, and so much more. If there is a fluorescent probe that can measure a specific characteristic, it can ...

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How To Use Flow Cytometry To Measure Apoptosis, Necrosis, and Autophagy

Written by Tim Bushnell, Ph.D “If one approaches a problem with order and method there should be no difficulty in solving it — none whatever.” — Hercule Poirot, Death in the Clouds Murder is a common theme in the mystery suspense genre. The detectives who solve these murders use a combination of observation and deduction to identify the guilty party. This metaphor suits measuring cell death. In biology, there are four major pathways for cell death. The study of the different ways cells die has become known as Cell Necrobiology, as coined by Darzynkiewicz and coworkers in their 1997 review article. Flow cytometry is ideally ...

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How To Prepare For A Flow Cytometry Experiment

Written by Tim Bushnell, Ph.D You never forget your first experiment. Usually, you remember all the things that went wrong that you need to correct for future experiments. In flow cytometry, there are so many things that you need to remember to bring to the first experiment, that having a checklist make sense. This checklist is divided into three phases — before the experiment, at the instrument, and after acquisition — with steps listed for each to help you perform your first flow cytometry experiment with ease. 1. Preparation before the Flow Cytometry experiment. Before embarking on the first flow experiment, there are several things that ...

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How To Differentiate T Cell State With Flow Cytometry

Written by Jennifer Snyder-Cappione, Ph.D. Have you ever wondered what has gotten into your T cells? Are they activated, resting, tolerant, senescent, anergic, exhausted, or maybe just having an off day? During short-lived immune responses, such as responses to acute infections or vaccines, there are three classically defined T cell states: naïve, activated memory, and resting memory. Here’s how to determine whether your T cells are naive, activated or resting memory, or exhausted, and how effector function plays into that. 1. Naïve cells. Naïve T cells have not encountered an antigen and secrete IL-2 and some chemokines. However, antigen-experienced (memory) T cells, have differentiated into effector cells ...

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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.

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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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Before taking this course, my experience with flow was limited to simple 2 color experiments. Then I was asked to perform a 10-color complete immunophenotype experiment in non-lymphoid tissues. With a ton of help from a long time co-worker and friend, I was successful with the effort. She highly recommended the Excyte Mastery Course to me and I can say she was right. This course truly elevated my knowledge and understanding of flow cytometry. It was very organized, concise and covered everything from understanding your instrument, optimizing it's performance, designing multicolor panels and executing a flow cytometry experiment with all the necessary controls. I look forward to learning more from the advanced seminars and webinars. I highly recommend this course.

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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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