Zeiss LSM 880, Operating Manual

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CHAPTER 1 - SYSTEM OPERATION
ZEISS Left Tool Area and Hardware Control Tools LSM 880
342 000000-2071-464 10/2014 V_01
The
PSF sampling
pull down menu allows you to
interpolate the result to different degrees. Since
the resulting PSF is an average of many PSFs
corresponding to the localized beads and every
bead is localized with the sub-pixel accuracy, one
can obtain after averaging a PSF with higher
resolution.
You can average the PSF by
1 (pixel size of the
resulting PSF is the same or
1
times as in the
original image), 3 (pixel size of the resulting PSF is
3
times smaller as in the original image),
5
(pixel
size of the resulting PSF is 5 times smaller as in the
original image) or 7 (pixel size of the resulting PSF
is 7 times smaller as in the original image)
times (Fig. 471).
The
3D PALM box will be automatically checked, if
the image data was acquired with the 3D-PALM
slider inserted. In this case the double lobe
structure of each bead will be aligned to the right
lobe.
In the
Background drop down menu you can
decide the way background is determined
(Fig. 472).
You have three options:
−
Local : Background is computed from the
periphery area of the mask defined in
Peak
Mask Size .
−
Histogramm : Background is calculkated
from the histogram as at 10% cut off value.
−
ROI
: Backgrund is compited from ROIs in the
image ( drawn with the Graphics tool). Note,
this option is only available if ROIs have been
defined in the image.
There are different filters that can be set by corresponding sliders or spin input boxes to select sub-
resolution beads.
Fig. 470 Experimental PSF – manual mode
Fig. 471 PSF sampling drop down menu
Fig. 472 Background drop down menu