Leica SL Confocal Instructions: Startup
Keck Microscopy Facility
Leica Confocal Instructions: Starting the system
Sign in on the log book
If the system is off, follow these instructions. Turn on only the lasers you need.
If the system is already running, check that the lasers you need are turned on. If not, go ahead and start them up using these instructions.
The switch (1) for the scan electronics is located below the key switches for the HeNe lasers.
Turn this on first to allow time for the scan head to initialize prior to starting the software.
The scan electronics must be on to use the Leica TCS confocal software.
Turning on the mercury lamp:
Turn on the mercury lamp at the power supply (2).
Wait for the lamp to ignite before proceeding (light will be visible in the lamp house).
The lamp should remain on for at least one hour. If turned off, it should cool for at least 20 minutes before turning on again.
ADJUST THE KNOBS ON THE MICROSCOPE LAMPHOUSE.
Ask Nathaniel for show you how to align the lamp if you think it's misaligned.
Turning on the laser(s):
Turn on only the laser(s) you need to use.
Argon Laser excites at 488 nm (GFP, Alexa 488, fluorescein (FITC), 458 nm (lucifer yellow, CFP) and 514 nm (YFP).
Use the following sequence when turning on the argon laser:
3) Turn on the large red switch. This starts the cooling fan which must be on before turning on the laser.
4) The LEVEL control on the laser should be at MIN (fully counterclockwise).
5) Briefly turn the key to START and then release to the ON position (like on a car). A yellow indicator light will come on.
6) Turn the LEVEL control to the position indicated by the green tape arrow.
GrHeNe Laser excites at 543 nm (dsRed, Alexa 543, Alexa 555, Alexa 568 and rhodamine). Note that Alexa594 and Texas Red are excited off-peak with this laser and will be dim.
7) Turn key to on (vertical). A yellow indicator light will come on.
HeNe Laser excites at 633 nm (Alexa 633, Alexa 647, Cy5, ToPro3).
8) Turn key to on (vertical). A yellow indicator light will come on.
If they're off, turn on the computer (9) and both monitors (10).
Turn on the microscope (switch (11) on right side as you face the microscope).
Open the valve on the top of the N2 cylinder (12). The pressure should read 60 PSI on the gauge (13). If there is no pressure, change the regulator (14) to the other cylinder (15) and let Nathaniel know.
The top of the anti-vibration table should rise slightly.
Contact Nathaniel Peters at firstname.lastname@example.org for additional information. Suggestions and comments are always welcome!
© 2015 Keck Microscopy Facility, University of Washington