LSM 510
ANNEX
LSM 510 META
Beam Path Configuration Guide for Fluorescence Microscopy ...
Carl Zeiss
03/06
B 45-0021 e
8-5
8.1.3
Multi Track Configuration
Multitracking is the method of choice for multi
fluorescence imaging. It has the advantage to
avoid artifacts based on emission crosstalk that
occurs when using simultaneous excitation and
detection. Laser lines are switched very fast and
channels recorded quasi-simultaneously.
The configuration of Multiple Tracks follows the
same rules described above for single track
configuration. The main difference is that each
track is configured to excite and detected only one
fluorescent dye to prevent cross talking (or two
dyes with non overlapping emission spectra).
•
Create a single track for both, Alexa 488 and
CY 3 detection separately (see above).
•
Open the Multi Track configuration window.
The system displays the Single Track setup as
track one.
•
Add a new track.
•
Click on track one, deactivate Ch 3 (red
emission detection) and switch off the green
laser line (561nm) in the
Line Active
check box
in the
Excitation
control window.
•
Click on track two, deactivate Ch 2 (green
emission detection) and switch the blue laser
line (488) off.
•
To extend the detection band for the green
light it is now reasonable to use the LP 505
instead of BP 500-550 in track one. This is now
possible since the green laser line is turned off
during detection of the green fluorescence
emission.
•
Use the Spectra window to check the proper
settings for each individual track as described
above.
Fig. 8-2
Configuration Control window
