11
Nikon Imaging Centre @ King’s College London
Z-stacks
Although not an ideal imaging mode for wide-field techniques due
to the presence of out-of-focus light, it is possible to perform Z-
stacks which may be used for 3D visualisation or for maximum
intensity projections. Select the
Z-stack
tab in the
ND Acquisition
window (Fig. 28).
1.
Press reset to remove any previously saved z-positions.
2.
Three options are available for setting Z-stack limits:
Top
and Bottom
(mark the limits of the stack),
Symmetric
(mark the centre of the stack) and
Asymmetric
(set the
number of slices above and below the centre).
3.
For the top-bottom option, whilst live imaging, adjust the
focus to find the top of your sample (use the Z value as a
guide) and click
Top
. Focus back through the sample to do
the same for the bottom plane of the sample. Stop imaging
to avoid sample bleaching.
Nikon
Imaging Centre
King’s College London
Fig. 28
Fig. 27
