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Flow cytometry: polychromatic or spectral?
Published on September 29, 2022 33 min
A selection of talks on Methods
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- Prof. John Fox
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Understanding statistics in epidemics and pandemics: lessons learned from COVID-19
- Prof. Sarah Ransdell
- Nova Southeastern University, USA
International biobanking: overview of key practices and policies
- Dr. Jim Vaught
- Editor in Chief, Biopreservation & Biobanking, USA
Data fusion: examples in fusing metabolomics and transcriptomics data
- Dr. Johan A. Westerhuis
- University of Amsterdam, The Netherlands
Hello. Today we are talking about flow cytometry in general, but more specifically on systematic differences between polychromatic flow cytometry and spectral flow cytometry, which has been a recent development. My name is Martin Buescher. I'm a physicist by training, but I got my PhD in Applied biotechnology. I'm running the BioPhysics group within the Miltenyi company that manufacturers flow cytometry devices and many other tools and systems around immunology and science in general.
First, I will briefly introduce flow cytometry from a historical point of view. After that's done, I'm going to talk about the fundamental principles that make up flow cytometry. From there, some practical consequences are derived that play a role in the use of flow cytometry. From there, we're going to discuss some practical consequences in real-life experiments, and from that, we'll deduce best practices in daily work with those devices. Then we are almost through and doing some final conclusions.
Introduction to flow cytometry. Flow cytometry was invented in the last century, in the 60s, mainly by Wolfgang Dittrich and Wolfgang Gohde. They published a paper on impulse fluorometry of single cells in suspension, where they used the light source, excited fluorescence, and worked out the fundamental principle of flow cytometry, which I'm going to talk about in a minute. Then in the 70s, this whole technology had developed quite a bit further and spear-headed by Leonard Herzenberg. They not only introduced cell sorting, and patented the technology by that, they also developed the systems further so that they could be used in routine labs. Then in the 80s, starting then until today, the systems have developed further and further and got more complex with more channels and are faster. Some bigger examples are publications from the mid-80s, where they did 8-color, 10-parameter flow cytometry experiments. By that, they could preserve a very different differentiated image of leukocyte heterogeneity in the blood. After that, things have developed further. Around the millennium, 17-color flow cytometry experiments have been conducted. More or less, the whole immune system, as it is known today, has been unraveled. Now the question is how to move from there and how to expand it further. This is probably with spectral flow cytometry.