CONFOCOR
3
Carl Zeiss
Options
ConfoCor 3
128 M60-1-0025
e
02/2010
(1)
Determination of the offset between LSM Scanner and FCS Stage Positioning
You can compensate manually or by using a Macro (only available for Rel. 2.x and 3.x versions).
Manual Adjustment
Do the manual adjustment according to the
following procedure:
•
Obtain an LSM image of a dried layer of a
suitable dye (Fig. 135). Optimize focus position.
To obtain the dye layer, dissolve the dye at a
concentration of 1 µM in ethanol, cover the
bottom of a suitable glass bottom chamber
with the dye solution, and let evaporate over
night in a laminar hood.
•
In the ConfoCor
Measure
tool select
LSM
Image
from the
Positions
panel. Mark the
points in the LSM image that you want to
bleach (Fig. 136).
•
Perform a pre-bleach with high laser power for
3 seconds. Close the pinhole to 10 in order to
avoid high count rates on the APDs.
•
Re-scan the image (Fig. 137). The bleached
spots should be visible.
Fig. 135
Image of dye layer
Fig. 136
Image of dye layer with selected
positions
Fig. 137
Image of dye layer after bleach
