CIAN Leica SP8 – short instructions, version 3.1, September 2015
page 6 of 10
3.2.5 - Acquisition Parameters
•
Format
, or number of pixels in the scan area; for
optimal format adjust to desired pixel size, see below
•
Scan Speed
; start with 400 to 600 Hz to avoid
limitations in zoom
•
Bidirectional X
scanning speeds up acquisition by
almost two-fold; may need to adjust phase
•
Zoom factor
can be adjusted in various ways:
o
Adjust slider
o
Enter specific factor
o
Activating “Zoom In” and drawing a region of
interest in the image (can be undone with
button that appears after use)
o
Using the zoom knob on the Control Panel
•
Display of
image and pixel size
; will change as
format and zoom are adjusted
•
Optimize xy Format
button to set the optimal number
of pixels to meet Nyquist criterion for best lateral
resolution, depends on NA of objective and emission
wavelength
•
Averaging
and/or
Accumulation
to remove noise
from high gain settings; note that accumulation sums
up the intensities of several passes and may lead to
saturation of the data space
•
Rotation
of the scan field by up to 100 degrees in
either direction; as with zoom, use slider, numerical
factor, or control panel knob
•
Panning
with the arrows positions the scan area
when zoomed in; middle round button to return to
center of field of view
•
Pinhole
will default to 1 Airy Unit based on the
objective NA and the emission wavelength defined in
the box (default 580nm). Change wavelength as
needed. Pinhole diameter can be adjusted with the
slider. Use Airy Units or diameter in
µ
m. Return to 1 Airy Unit by pressing the “Airy 1” button.
3.2.6 Acquisition modes besides z series
a) Time series
Select acquisition mode that includes time (t), set time
interval (for no delay between images, check
“minimize”), define parameter for duration.
b) XZ scan with SuperZ galvo stage
Select xzy as acquisition mode, move white line to desired scan position, start live scan; adjust z
position with SmartMove or focus knob, use right-most control panel knob for y position.
When switching back to xyz mode, the white line can be moved to the future z position.
c) Spectral scan
Select acquisition mode that includes lambda dimension; set up laser(s) and detector(s) to be
used, top and bottom wavelength, interval, emission band, and/or number of steps.