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ELYRA 7
Left Tool Area and Hardware Control Tools
ZEISS
03/2019 V_02
000000-2262-999
69
Please note that the number of cycles in
Time Series
must be an integer multiple of the
Sequence Size
. If
changing the number of
Sequence Size
in a way that
the cycles in
Time Series
is not any more an integer
multiple of
Sequence Size
, the software will
automatically update the number of cycles of
Time
Series
to the next integer multiple. E.g. if the
Sequence Size
was 100 and the cycle number set to
10000 and now the
Sequence Size
is changed to 90,
the next integer multiple of 10080 will be selected
(Fig. 85).
After having defined your track configuration you can
set the acquisition parameters in the
Channel
tab
(Fig. 5). You can select for each track independent the
laser lines used and their power settings. When
setting
Exposure Time
,
EMCCD Gain
,
TIRF Angle
or
Collimator Position(s)
(the latter only visible in
Show all
mode) for one track, the other track will be
automatically updated to the chosen settings. These
parameters can therefore not be set individually for
each track.
Note, that the
EPI
and
TIRF
button usage (pressed or
depressed) will not be updated, but the values will. E.
g. if in every track in
TIRF Angle
the button
EPI
was
pressed and now in one track the
TIRF
button is
pressed, only in this track the button usage is
indicated. In all other tracks
EPI
stays selected,
however, the last change will overrule all previous
settings and each track will have TIRF illumination.
In the
Online Processing Options
tab you have the
possibility to activate Online processing, however,
Automatic Laser Control
and
Holding Focus
will
not be available for sequential laser switching and
their functions are greyed out.
Once, the settings are made and the time series
defined, press the
Start Experiment
button. One
channel will be open for each track (Fig. 86) and
sequentially filled with the images, whereby channels
are switched every number of frames as defined in
Sequential Size
.
Frames for each channel are stitched together and
treated as a continuous run, therefore absolute time
information will be lost for each channel. The image
will be treated as a two channel SMLM image and can
be processed accordingly (Fig. 87).
Fig. 85
Cycles of "Time Series" set to
10 080 after "Sequence size" has
been changed from 100 to 90
Fig. 86
Track acquisition settings in the
"Channels" tab