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ELYRA 7
Left Tool Area and Hardware Control Tools
ZEISS
03/2019 V_02
000000-2262-999
273
5.4.3
Maintain tool group – Adjust PALM Slider
Open the
Adjust PALM Slider
tab to position the 3D-PALM slider at the correct position (Fig. 461).
You can select the illumination scheme by pressing the corresponding
EPI
(Epifluorescence),
HILO
(high
inclined and laminated optical sheet) or
TIRF
(Total Internal Reflection Fluorescence) button. The selected
button is highlighted.
With the
Position
slider or the
Position
input box, you can set the position of the 3D-PALM slider.
You can save the position for the objective in use by pressing the
Store
button, or for all objectives by
pressing the
Store for all objectives button
. The saved positions will be used, whenever the respective
objective is in use.
There are three ways to determine the optimal position of the slider (3D slider in active position):
−
HILO
illumination using 100 nm bead sample. In this case, focus on the beads (the position where
the gap between the two lobes points in a 45
O
direction) and increase laserpower or camera gain
to slightly oversaturate judging with Range Indicator. Position the slider so that both lobes become
the same intensity (Fig. 462).
Fig. 461
Adjustment window for the 3D-PALM slider
Fig. 462
Adjusting the 3D-PALM slider in HILO using equal
illumination intensities of beads with non-optimal (left
panel) and correct (right panel) positions.