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Rolly Wiegand – CALM Leica SP5 manual
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1.7. The software will start the initialisation of the microscope stand and the scan head,
and it cannot be used during this process. After a few minutes, the message shown in
Fig. 2B will appear, asking whether you want to initialise the motorised stage.
Fig. 2B
You should have already checked that the objectives and the condenser are out of the way
of any part of the stage (see 1.5., above). If you choose ‘Yes’, this will initialise the
motorised stage, which is essential if you want to use stage position coordinates (‘Mark
and Find’, ‘Tile Scan’). If you are restarting the software after a crash and you do not want
to loose the previous positions of your sample, click ‘No’.
Wait until the system is completely initialised.
1.8. Selection of the objective lens
The LAS AF software will open showing the ‘Acquisition’ page. Select the objective you
want to use in the ‘Beam Path’ window (see Fig. 3).
IMPORTANT
: Never change the
objective by manually turning the turret itself!
Fig. 3
The current objective lens is shown in the light path diagram of the window as seen in
Fig. 3. Before you start, you must check the immersion medium specifications of the lens
you want to use. This information is given in the ‘Beam Path Settings’ (Fig. 3), but you can
retrieve more detailed information about each lens from the objective selector by clicking
on the ‘Objective …’ in the configuration tab (Fig. 4, see circle). This will pull out the
following list as shown in Fig. 4 with all the lenses available on the system, including the
immersion medium specifications. Click the objective you want to use and apply the
appropriate, correct immersion medium, which could be:
DRY – air (never apply any fluid to theses lenses)
OIL – immersion oil only (use the provided Zeiss immersion oil)
WATER – water only (MQ water)
