- Ensure the microscope has been switched on and initialised, and Zen Black software has been opened for at least 3 hours prior to imaging.
- It is advisable to use the 100x TIRF lens to do PALM/STORM although this is not strictly necessary. Imaging in TIRF reduces the background and makes PALM reconstruction easier, however TIRF only illuminates objects very close to the coverslip.
- When ready to image, select laser WF option in the light path menu. Select 'TV1', which is the Andor CCD for PALM/STORM imaging. It is advisable to have the 1.6 optovar selected too.
- Set up a track for your most easily located channel and look for the right focus position in Epi mode.
- Then change to TIRF and move the tirf adjustor slider slightly until TIRF is achieved. This is usually marked by a sudden significant decrease in background fluorescence.
- Reduce image size if required.
- Set up a time lapse for ~10000-30000 images.
- Image
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