Recently, I am trying to conduct a FRAP experiment on coacervates using a Nikon Eclipse ti2 Confocal Microscope. I notice that in many of the papers, the stimulation can be conducted in a very short period of time and the coacervates can be photobleached effectively. However, when I use the ti2 model, the photobleaching is not strong that it takes a longer stimulation time (~5-6 s) to reduce the ROI intensity by half (in some cases not even half). I consulted the technician and he suggested that it is the problem with the model, stating that the earlier model was better at doing FRAP and the new model has sacrificed the FRAP part for better imaging. Also, it costed some money to upgrade the laser for better FRAP.

I am wondering if anyone has experienced similar problem before using the same model.