Zeiss LSM 880, Operating Manual

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CHAPTER 5 - MULTIPHOTON LASER SCANNING …
ZEISS Non Descanned Detection (NDD) LSM 880
16 000000-2071-464 10/2014 V_01
2.4 Filter for non descanned
detectors (NDDs)
To make the best use of NDDs it is necessary to
achieve optimal blocking of the excitation laser
light. This is done in the same way for all
LSM systems with NDDs independent of the
microscope used.
The first NDD module, which is attached to the
NDD port, is either a two channel module
(Fig. 10/
1
) or a BiG.2 module. Inside those modules
right at the opening towards the microscope is a
small clamping that holds a blocking filter (1442-
160 or 1442-161; order# Feb. 08). Both filters are
short pass filters which block the IR light of the
multiphoton laser with an optical density (OD) of at
least 7. If none of these filters is installed, the
image quality will not be sufficient as too much
laser light will get to the detectors. Always make
sure on of these filters is in place.
To install them, open the metallic grey lid of the NDD module turning the thumbscrews (Fig. 10/ 1 ). Once
the lid is removed the clamping is accessible. Slid in the filter, which is attached to a small holder, into the
clamping with the little screw on the side of the holder situated on the left side of the holder when
facing towards the microscope.
All emission filters offered for the NDD modules have an additional IR blocking capacity of OD 5. Other
filters without these characteristics may lead to inferior image quality.
All secondary dichroic filters for the NDDs Push&Click filter cubes are long pass filters. They split the
emitted fluorescence according to the indicated wavelength into the longer and shorter spectral range. In
this respect the dichroic filters are specified in the spectral range of 380 to 800 nm.
The Emission filters are mounted into Push&Click filter cubes which are clamped into the slider of the
NDD modules. NDD modules with two channels hold two Push&Click filter cubes. NDD modules with one
channel hold one Push&Click filter cube. The NDD module to be used for the end of the cascade does
not have a slider for a Push&Click filter cube.

To change the cubes simply take out the sliders which hold the cubes by unscrewing them from the
rest of the module using the thumbscrews (Fig. 10/
1
).

The filter cubes can now be mounted onto the slider by sliding the side noses of the cubes under the
corresponding spring latches. The orientation of the dichroic filters and emission filters is indicated on
the slider.
The arrangement of the filters inside the NDD modules depends on the orientation of the modules on the
microscope.
For LSM NLO systems on Axio Observer the NDDs are attached to the side port (up to 5 channels are
possible) or the transmission port (maximum of two channels is possible). All dichroic filters offered by
ZEISS for this modules, are long pass dichroic filters. Therefore the blue part of the spectrum is detected
with the first NDD whereas the red part is guided further on to the following NDDs (Fig. 11).
Fig. 10 Thumbscrews of the P&C sliders