ZEISS
Left Tool Area and Hardware Control Tools
ELYRA 7
194
000000-2262-999
03/2019 V_02
If the
discard overlapping
molecules
was
selected from the
Settings
drop down menu, all
identified peaks with overlapping circles will be
indicated in red and discarded (Fig. 316). If
ignore
overlap
or
account for overlap
was selected,
overlapping peaks will not be indicated and all
peaks will be circled in white.
−
If
discard overlapping molecules
is
selected, all overlapping peaks, indicated by
red circles will be discarded. This will result in
the highest accuracy, which is defined as the
deviation of the fitted to the real peak
position and localization precision (the
standard deviation from the fit) (Fig. 317,
A
).
This method is therefore the method of
choice for single emitters and little overlap.
−
If
ignore overlap
is selected, all single and
overlapping peaks will be taken into
account. However, fit accuracy will be
decreased when multi-emitter events are
fitted as the distribution of one emitter will
influence the distribution of an overlapping
one without this fact taken into account.
Hence accuracy might suffer for multi-
emitter events whereas localization precision
will be sustained (Fig. 317,
B
). This method is
most suitable for slightly overlapping
molecules.
−
If
account for overlap
is selected all single
and multi-emitter events will be taken into
account. For multi-emitters the overlap of
neighboring molecules is taken into account
so losses in accuracy are minimized. There
will be a slight decrease in the localization
precision (Fig. 317,
C
). This method should
be used, when molecules heavily overlap.
Fig. 316
Preview window of the image
container for SMLM processing with the
discard molecules (A), ignore overlap
(B) and account for overlap (C) selected
Fig. 317
Analyzed data set of Fig. 316 using discard overlapping molecules (A), ignore
overlap (B) and account for overlap (C).