Scanning An Image - Zeiss LSM 510 Manual

Scanning an image

• Click on the Scan button in the Acquire subordinate toolbar to open the Scan
Control window.
The Scan Control window appears (Fig. 16).
Setting the parameters for scanning
• Select Mode in the Scan Control window.
• Select the Frame Size as predefined number of
pixels or enter your own values (e.g. 300 x 600)
in the Objective Lens, Image Size & Line
Step Factor panel. Click on the Optimal
button for calculation of appropriate number of
pixels depending on N.A. and λ.
The number of pixels influences the image
resolution!
Note: When using an Axioskop 2 FS
the objective that is in use in the Scan Control
window. This ensures correct calculation of
pinhole, Z stack optimization etc.
Adjusting the scan speed
• Use the slider in the Speed panel (Fig. 16) to adjust the scan speed.
A higher speed with averaging results in the best signal to noise ratio. Scan speed 8 usually produces
good results. Us speed 6 or 7 for superior images.
Choosing the dynamic range
• Select the dynamic range 8 or 12 Bit (per pixel) in the Pixel Depth, Scan Direction & Scan Average
panel (Fig. 16).
8 Bit will give 256 gray levels, 12 Bit will give 4096 levels. Publication quality images should be acquired
using 12 Bit data depth. 12 Bit is also recommended when doing quantitative measurements or when
imaging low fluorescence intensities.
Setting the scan averaging
Averaging improves the image by increasing the signal : noise ratio. It can be achieved line by line or
frame by frame. Frame averaging helps reduce photobleaching, but does not give quite such a smooth
image.
• Select the Line or Frame mode for averaging.
• Select the desired scan average method Mean or Sum in the Method selection box.
If you are using the Mean method, the image information is generated by adding up all scans pixel by
pixel and then calculating the mean value.
03/06
, indicate
MOT
Fig. 16 Scan Control window, Mode settings
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