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Lightsheet Z.1
Center Screen Area / Image Containers - Display and Image Analysis
Carl Zeiss
02/2013
000000-1790-528
225
Histogram Threshold:
When the
Threshold
tick box is selected, the
controls to set a lower and upper threshold for the
histogram calculation are displayed (Fig. 337). The
threshold can be set for each channel (channel
selection pull down menu).
4.10
Colocalization View
The
Colocalization
function permits interactive
analysis of two channels of an image by computing
a scatter diagram (co-localization).
The settings of the
Dimensions
,
Display
,
Player
,
and
Graphics
view options control blocks apply.
The additional view-specific
Colocalization View
Option
control block is shown in Fig. 338.
Any changes done with this View Option control
block are effective immediately.
The Image Display in the Colocalization View shows 3 panels: the scatter diagram, the data table, and the
pseudo-colored image display (see Fig. 339 and Fig. 340).
Colocalization is defined by the presence of two or more different molecules at the same location
in a specimen. However, in the context of digital imaging, the term colocalization refers to colors
emitted by fluorescent molecules detected by the same pixel in the image. It is important to be
aware of the fact that colocalization cannot be analyzed for fluorophores with similar emission
spectra. Accurate colocalization analysis is only possible if the fluorescence emission spectra are
well separated between fluorophores and the correct filter sets are used for data acquisition. If
spectral bleed-through artifacts are present because of spectral overlap between the fluorophore
emission spectra, or due to the use of incorrect filter sets, colocalization measurements will be
meaningless. To avoid this, the fluorophores must be carefully chosen and matched to the
excitation laser lines to obtain the maximum excitation efficiency while still maintaining a useful
degree of separation between emission wavelengths. The choice of fluorophores is crucial for
colocalization analysis.
Fig. 337
Histo View Option control block –
Threshold
Fig. 338
Colocalization View Option control
block
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