Hi there. Can DAPI be excited with 440 nm wavelength? Maybe still a bit of tail is there from the absorption spectrum, yet we are much in the emission spectrum already. Making me think what we get is mainly just stimulated emission. Anyone has experience on a similar test?
Hey Marco Salerno! Exciting DAPI with a 440 nm wavelength is a bit unconventional, but it's not entirely out of the realm of possibility. While DAPI's absorption peak is around 358 nm, there can still be some excitation with a 440 nm wavelength due to its broad absorption spectrum. However, you're right to suspect that what you're mainly observing is stimulated emission rather than true excitation.
If you're considering this approach, it's worth conducting some test runs to see what kind of results you Marco Salerno get. Keep in mind that while you Marco Salerno might see some fluorescence, it may not be as robust or specific as when using the optimal excitation wavelength. Additionally, be mindful of potential photobleaching and phototoxicity effects at higher wavelengths.
As for similar tests, there might be some scattered experiences out there, but it's not a commonly explored approach. If you Marco Salerno do proceed with it, documenting your methodology and results could contribute valuable insights to the scientific community. Always an adventure to push the boundaries! Looking forward to hear from you about your results.
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