I am using stimulated emission based pump-probe microscopy technique to detect fluorescence. For application purpose, I want to do some cell imaging. But, When I use excitation beam at 635 nm to the fixed cell labeled with ATTO647N dye I do not get any fluorescence signal by using photodetector (PDA36A) or PMT. I do not understand where is the problem.
Can you elaborate what is your "stimulated emission based pump-probe microscopy"?
I suggest you just forget the cell, check the dye. If that works, it means you have something wrong with your labeling procedure.
Please visit this link below for more information
https://www.osapublishing.org/oe/abstract.cfm?uri=oe-20-10-11445
I have been using ATTO647N dye. It works very well in my setup. Once I use fixed cell labeling with dye I can not detect any signal in the detector. I do not think it is labeling problem or detector problem. It could be because of low concentration but I have found in many papers they used this technique to image cell and tissue.
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