Zeiss LSM 880, Operating Manual

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CHAPTER 1 - SYSTEM OPERATION
LSM 880 Left Tool Area and Hardware Control Tools ZEISS
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The
Lasers
functionalities for laser control
are also available in the Acquisition tab
(Fig. 268). Only that all visible and
invisible lasers are displayed together (als
applies if
Show All mode is not set in the
Light Path
tool window).
Procedure for setting the laser power
•
Set the laser attenuation using the sliders in Lasers from the Laser control panel in the Light Path
tool or
Acquisition
tool. Open the
Count Rate
window by clicking the
Count rate
action button in
the FCS tab. A good starting point is to set the intensity in such a way that a count rate between 50
kHz and 200 kHz is obtained.
•
For most dyes, the Counts / Molecule setting should be optimized in a second step to a value just
under its maximum by changing the laser power. If carriers of different slide thickness are employed
the Counts / Molecule setting should be optimized by using the correction ring on the lens. The
correction ring is turned counterclockwise or clockwise until a maximum value is obtained. The
correction ring should also be used for adjusting the Counts / Molecule setting whenever the
immersion media is changed. This is especially important in cases where the refractive index of the
immersion media is different from that of the sample. When finished, close the
Count Rate window
using the
Close
button.
Procedure for defining a new beam path configuration
•
First tick the laser or lasers (in case of cross-correlation) you need for excitation of your sample.
•
Then choose a major dichroic beam splitter which should corresponds to the chosen laser(s).
Please note that you can use IR lasers for FCS, but there are no specifications for the performance
of the instrument for this excitation.
•
Activate the channels you want to use. It is recommended to give the channel a name which is easy to
remember.
•
Set the laser power.
•
Perform a pinhole alignment or adjust the collimator settings.
•
Store beam path or method, which uses the beam path.
BiG.2 as 2 Channel Module or NDD you have to choose the right SBS and EF in the filter cube
and place it manually. The filters are indicated to the software within the Filters Tool window
(Maintain tab). Otherwise the system does not display the spectral range of the filters.
Fig. 268 Laser lines in the Acquisition tab