Leica Sp5 WLL STED, Tutorial Manual

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14
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By using the depletion laser 592nm, the samples can be bleach very quickly, so first scan the sample in
standard Confocal mode to find the focus and prepare the STED configuration close to work.

Set the WLL laser line that you need to use ( 10 to 30 % ).
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Select the Hyd 2 detector and set it at 100%.
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Introduce to the Hyd a bandwidth that is 10nm fahter
to the WLL line that you set, and maximum 10nm lower
than the wavelength of the laser depletion ( +/-582nm).

Use a format like 512x512 or 1024x1024 .

Use a scan speed like 400HRZ .

Use a zoom factor from 4 to 6 .
STED configuration ( step1)