This webinar covers the basic components of a flow cytometer, how to interpret a dye excitation/emission spectrum, how data is displayed, basic gating demonstration, and common statistics and terminology used in flow cytometry.
Thanks for watching and thanks for your question. You can find Flow Cytometry, Fluorescence, and Imaging Basics in our Molecular Probes School of Fluorescence - www.thermofisher.com/us/en/home/life-science/cell-analysis/cell-analysis-learning-center/molecular-probes-school-of-fluorescence.html
We contacted the team that created the deck, if it is still available, we will reach out to you to let you know how you can get a copy of it. Thanks for the request.
Can we apply two different lazers (e.g., FITC and PE) at the same time to excite cells, and then detect and sort only cells that are exicted by both lazers, and get data from computer? This would be the same effect by Image J. (e.g. Getting FITC and PE images, respectively from fluorescence microscope and then merge them, which generates yellow pseudo color)
Hi Elle. Thanks for your question. Histograms are used as a way to look at a “shift” in a single channel (antibody). They’re not as informative as a scatter plot, where you would be able to look at he overlap/co-indicence between two different antibodies. For additional technical support, please contact us at thermofisher.com/askaquestion. Thank you!
Hi Chiedu, if you're looking for more videos or webinars on flow cytometry, we have a whole page full of them at the link below: www.thermofisher.com/us/en/home/life-science/cell-analysis/flow-cytometry/flow-cytometry-learning-center/flow-cytometry-resource-library/flow-cytometry-educational-videos-webinars.html
The answer to question #1 fluorescence emission is always longer than excitation should be false. In cases of multi-photon, energy upconversion for example, excitation wavelength is longer than emission. I guess those 4% people answered correctly are those from physics or photonics or laser application background like myself :)
Of course... but this video assumed a regular "single photon" laser excitation behavior. And even in this very context, if you have a closer look to the different spectra of emission of your fluorochromes, you will see that some photons do have a shorter emission wavelength than the laser used to excite them... typically FITC photons below 488nm... In reality, these photons exist in a "pre-excited state" before they receive the energy (h.v) from the laser. You can read Shapiro to know more about it.
