I have a TIRF setup with a 488nm laser. I am using a quad band dichroic (405/488/561/635) in combination with a quad band fluorescence filter (446/523/600/677). When the objective is not present in the light path no laser is being transmitted through the dichroic and is perfectly reflected. However, the moment I put the TIRF objective, I can clearly see the laser leaking through to the camera. I have tried -
1. Changing the filter sets.
2. Changing the objective.
3. Checked the alignment.
Nireekshit Addanki Tirumala
Thanks for your responses. The problem was the dichroic cube that was not fitting in properly, i.e the angle was slightly off from 45.
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