While operating the LAS X software for 4 color, my HYD 3 detector intermittently overloaded, with the same settings. I made sure the absorbance spectra is separate from the laser spectra. I am not sure why it is happening, could it be due excessive luniniscence from 'RFP' that is being detected. I also see some noise now, comet effect when going live, does that mean my HyD detector need to be replaced?
Hi
I hope your sp8 has aobs( aqusto optical beam splitter) ,not spinning disk, so u will have levarage of using any detector without restriction,
We also have same problem,
Hyd goes off due to overloading
I can suggest
1.use lower gain
2.use standard setting for acquiring not photon count
3.use detector in slightly large time gap because it will take time to normalize ,
4.put emission maxima in 1au for individual flurophore, by default it is 580 nm and many times people mistake it being adjusted to 1 airy unit,as defraction pattern of each light wavelength will be different
5.Apart from all you can look at negative control samples if you r still getting fluorescence then I will go for pmt only ,i guess you will have two pmt so you can use pmt 1212 sequentially to carry the acquisition,
Thank you for the answer, I would definitely look into the suggestion, I am trying to fiddle with using larger time gap as of now. I have been using PMT since, for everything except far-red.
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