I am imaging some optic nerve head sections using non linear microscopy (SHG/TPEF simultanously). I am going to merge the two channels (SGH green and TPEF red) in order to see the co-localisation of collagen and elastin. I am getting some autofluorescence from TPEF coming from the whole sample so it makes difficult to distinghuish elastin from everything else.
What can I do to avoi/remove/reduce auto fluorescence?
Thank you