If you intend to use flow cytometry on a regular basis, it is a good idea to do some reading on how fluorochromes are actually excited by a laser and where their emission spectra lies. Once you have an understanding of this it is very easy to design your stain sets without overlapping emission spectra. For now you could try utilizing an online generator that can help you narrow down your fluorochrome choices (http://www.biolegend.com/spectraanalyzer). However, I must stress that it is essential for you to gain an understanding of the underlying mechanism that determines how to choose fluorochromes that will not have overlapping spectra.
If you intend to use flow cytometry on a regular basis, it is a good idea to do some reading on how fluorochromes are actually excited by a laser and where their emission spectra lies. Once you have an understanding of this it is very easy to design your stain sets without overlapping emission spectra. For now you could try utilizing an online generator that can help you narrow down your fluorochrome choices (http://www.biolegend.com/spectraanalyzer). However, I must stress that it is essential for you to gain an understanding of the underlying mechanism that determines how to choose fluorochromes that will not have overlapping spectra.
Robby and Amanda are quite right. We could guess what would work on your system and in your panel but quite a few factors come into it.
Low expressed markers should be on bright fluors and conversely Highly expressed markers should be on weaker fluors (i.e. CD3 on PacBlue and CD34 on APC)
if filters are set up differently with your detectors at your facility it may not work properly
As has been said before, depends on the particular instrument that you have and the configuration of your instrument. Once said that, there are some tools that might help you to design your panel such as fluorish or chromocyte (links below). The advantage of chromocyte is that it also provide useful information for newbies in cytometry that you might find helpful. Also, there is an international effort to validate what has been called optimized markers for immunophenotyping panels (OMIPS) in cytometry (ISAC) which you can access (link to OMIPS). Best regards.
http://onlinelibrary.wiley.com/journal/10.1002/%28ISSN%291552-4930/homepage/information_on_omips.htm
http://www.fluorish.com/
https://www.chromocyte.com/
I really like BDBiosciences Spectrum Viewer, but an even more in-depth option is the free program available at Fluorish.com. It helps you create a panel based on the configuration of the instrument you use and has fluors available from many different manufacturers that may be more affordable. Once you've selected an antibody (say CD4-FITC), when you go to select your next cell marker, it will no longer show you options for fluorochromes that would interfere with the ones you have already added to your list. It also has direct links to purchase.
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