Zeiss LSM 880, Operating Manual

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CHAPTER 5 - MULTIPHOTON LASER SCANNING …
ZEISS NLO (Non Linear Optics) Laser for LSM LSM 880
4 000000-2071-464 10/2014 V_01
1.2 Imaging Setup and Channels tool
•
Setting the wavelength of the laser/OPO
•
The wavelength of the laser and OPO can be set in the Imaging Setup tool in the pop up window of
the laser icon or in the Channels tool. Enter the desired wavelength for the laser and the OPO in the
input box. The OPO can only be pumped at certain laser wavelengths typically between 780 and
850 nm. This is dependent on the type of laser used for pumping the OPO. Refer to the manual of the
OPO for further information. The software shows the status of the laser and the OPO in the
Laser
control tool (see section 1.1)
•
The laser intensity (transmission through an Acousto-Optical Modulator, AOM) is controlled in the
laser control pop up from the
Imaging Setup
tool or in the
Channels
tool. The intensity of the OPO
is controlled via a manual neutral density filter. Changing the intensity is possible via a slider in the
Imaging Setup
tool or the
Channels
tool.
Note: With the ND filter as attenuator it is not possible to perform a line or frame fast Multitrack or
to bleach arbitrary ROIs other than rectangular regions.
Application of the NLO laser requires special main dichroic beam splitters to be activated in the
Imaging
Setup tool.
NLO-excited signals can be detected in every channel. However, it is important to efficiently block NIR
(near infrared) excitation stray light from the detector.
The following filters are designed for detection of NLO-excited fluorescence signals:
−
MBS 690+: main dichroic beam splitter reflecting NIR excitation at 690 nm and above, transmitting
shorter wavelengths
−
MBS 750+: main dichroic beam splitter reflecting NIR excitation at 750 nm and above, transmitting
shorter wavelengths
−
MBS -405/760+: main dichroic beam splitter reflecting NIR excitation longer than 760 nm,
transmitting shorter wavelengths and reflecting 405 nm excitation for simultaneous detection of
signals from NIR and 405 excited fluorochromes.