Troubleshooting the 2p rig
If you cannot get an image on the 2-photon, check the following things:
Check the laser performance: under the "2P laser" tab on PrairieView, examine how much power the laser is producing in mW (should be several 100mW) and whether the 2P laser shutter is open or closed (should be open). If you don't have power, check with Arani or Steve.
Check the path items:
Is the 2P/camera rod in the out position?
Is the turret in position 1? Does the turret appear to be solidly in position (for example, if you look under epifluorescence, do you see things fine?)?
Is the Pockels cell power on, and is the bias dial set to the same number as listed in the laser power configuration?
Is the Pockels cell turned up a bit on the PrairieView software? (Should be yes)
Is the PMT amplifier power on (a "rocker switch" on the right of the amplifier, which is the box in the rack immediately beneath the computer)? Is at least one of the PMTs turned up to about 800-900 on the software? If you turn up the PMT gain to 1250 and scan, do you see noise that looks a bit like "snow"? If you don't see snow, there is a problem with the PMT, possibly that the amplifier is turned off.
Does the power meter that sits on top of the laser indicate there is laser light on the table? (Should be yes)
When you start/stop scanning, do you hear the click of the Prairie shutter in the back of the microscope going open (start scanning) and shut (stop scanning)? Should be yes. If not, check to make sure that either 1) the screen is pulled all the way down so that the "buttons" on the front of the box are pressed, or 2) the override magnets are in place. Otherwise, check with Arani or Steve, the cable might be bad or unplugged.