Zeiss LSM780
Start up procedure for LSM 780
1) Turn on Main switch and the two switches labelled ‘systems/PC’ and
‘components (beside the keyboard)
2) Turn on the black switch on the argon laser power supply (to the left of the
microscope on top of the real time PC)
3) Turn the key on the argon laser power supply to ‘on’ (you will hear the fan
start)
4) Turn on the HXP 120 (on top of the white laser module)
5) Turn on computer
6) After ~5 mins flick the small silver switch on the hand controller for the
argon laser to run (the switch is located on top of the laser module near
the power supply for the argon laser). If this is done to early, the laser will
start to click; flick the switch back to standby and try again in a couple of
minutes.
7) Wait for the green light to appear on the hand controller for the argon
laser. If the red light appears, turn the dial on the controller anticlockwise
until the red light goes out.
8) Log onto the system as ‘LSM user’ (no password required)
9) Start ZEN 2010 software and press ‘start system’
10) To toggle between using the microscope for imaging and viewing
fluorescence/brightfield through the eye pieces use the ‘ocular’ and ‘LSM’
tabs in the top left hand corner of the software. Remember to press
‘online’ in the ocular menu if you want to see your sample through the
eyepieces.
11) Use the hand pad of the microscope to change objectives, filter blocks and
shutters under ‘control’ and the sub-menus ‘objectives’ and ‘reflector’
12) Remember the epifluorescent light source (the HXP 120) is controllable
using the dial on the front. Decrease the intensity if your sample is likely to
bleach.
Bioimaging @Rothamsted Research
Zeiss LSM780
13) Once sample is located and in focus, move to the ‘LSM’ tab in the top left
hand corner and then go to ‘Smart Setup’ to configure the system
14) ‘Autoexposure’ will then produce an image on the screen. Focus the
sample using ‘live’ and then change the Master Gain and Digital offset
values under the ‘channels’ drop down menu to balance the image. Use
the look up table short cut (the coloured square beneath the image) in
order to set the levels correctly.
15) The pinhole will not automatically default to 1AU. If you want to image
your sample with optimal pinhole size press 1AU in the channels menu for
each track you are using. Reset the master gain and digital offset values
until you have the image you require
16) Acquisition parameters such as resolution and line average can be found
under the ‘Acquisition menu’
17) Zoom etc can also be found in the acquisition menu under Scan Area. The
crop function is below the image window
18) ‘Reuse’ is at the bottom of the image window; open up a previously taken
LSM file and press reuse to load up the settings used to acquire that
image.
Shut down procedure for the LSM 780
1) Turn key on argon laser power supply to ‘0’. Allow to cool for ~5 mins until
the fan cuts out (you will hear the noise stop). Once fan has stopped, turn
off black button and flick the silver switch on the hand controller to
standby.
2) Turn off the HXP 120
3) Turn off computer in the usual way
4) Turn off the 3 switches on the control unit under the monitor
5) Clean all objectives and replace dust cover
Bioimaging @Rothamsted Research