Leica TCS SP8 SMD User Manual page 185

For fcs, flim and flcs
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Reversed image with inverted microscope
Please note that with an inverted microscope the image is inverted. In
this case, the bottom reflection (between glass and specimen) is the
important signal that has to be optimized.
6. Go to a higher zoom (e.g. 4 to 8) and activate a continuous xz scanning operation with the
Live image acquisition button.
7. Rotate the correction ring into a position that provides the highest contrast in the
reflection between the cover slip and the specimen (greatest possible intensity and
symmetry, narrowest possible intensity maximum without secondary maximum values;
Figure 162 and Figure 163).
Figure 162:
Reflection between cover slip and measurement medium with poorly set position of the
correction ring: The image is out of focus, several secondary peaks are visible
Figure 163:
Reflection between cover slip and measurement medium with correctly set position of the
correction ring: The image is sharp, and secondary peaks are scarcely visible
F(L)CS Data Acquisition
185

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