I am using a BD FACSMelody which currently has three lasers and 8 channels. When designing panels, I'm concerned about the spectral overlap between different fluorophores in some of the channels. Can anyone give me their perspectives on having used these fluorophores and whether they can use them in conjunction with each other? I tend to avoid using PE-Cy7 and APC-Cy7 together because, even though they should emit slightly differently, I always have trouble separating the signals. Is this common?
Yes, it is common to experience difficulty in separating signals when using PE-Cy7 and APC-Cy7 together. This is because they have similar emission spectra and can overlap with each other. To reduce spectral overlap, it is important to use fluorophores with distinct emission spectra, such as PE-Texas Red and APC-Cy7. Additionally, it is important to make sure that the lasers are properly aligned and that the correct filters are being used. Sometimes, a combination of filters and lasers can help improve the separation of the signals.
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