Performance Features Of The Lsm 880 - Zeiss LSM 880 Operating Manual

CHAPTER 1 - SYSTEM OPERATION
ZEISS Hardware Aspects LSM 880
2.5

Performance Features of the LSM 880

Optical and Mechanical Aspects
The highly integrated system design makes for the shortest possible optical paths, top-grade optical
precision and high stability. The Airyscan module can be mounted to the LSM 880 scan head of any
configuration and with upright or inververted microscopes. The BiG.2 module is either directly attached
to the scan head and can be used in combination with upright and inverse microscope stands.
The spectral range available extends from the UV to the IR region.
For the VIS (visible-light) Laser Module, the user can select from up to six lasers with wavelengths of 633,
594, 561, 543, 514, 488, 458 and 405 nm. The UV Laser Module provides wavelengths of 355 nm. Ti:Sa
Laser provide pulsed laser light from 680 to 1300 nm for Multiphoton imaging (NLO). UV and visible
lasers are coupled to the scan head via polarization-preserving single-mode optical fibers. Ti:Sa lasers are
coupled to the sytem via free beam coupling kits. One variable beam collimator each for the UV, NLO and
visible ranges provides optimum adaptation of the respective laser wavelength to the objective used and,
thus, optimum correction for Z aberrations.
Acousto-optical tunable filters (AOTF) adjust the necessary brightness for up to 8 laser lines within
microseconds. Ti:Sa lasers are attenuated via AOM within less than a microsecond.
The internal image acquisition channels usable for reflection or fluorescence, and an additional
transmitted-light channel are ideal for the investigation of multiple fluorescence specimens. Airyscan and
BiG.2 are attached directly to the scan head. Both are equipped with Gallenium Arsenide Phosphide
detectors (GaAsP). The diameter of the pinhole and the XY positions can be optimized, and the desired
emission can be selected. This adjustment also includes positioning along Z. For the simultaneous
registration of multiple fluorescence signals, identical optical sections can be obtained in each confocal
channel.
The microscope's transmitted-light channel is equipped with a photomultiplier, too. It is therefore
possible to superimpose a multiple fluorescence image on a brightfield, differential interference or phase
image.
Airyscan is equipped with a filter wheel consisting of 8 positions of which 6 are used for specific double
band pass emission filters. BiG.2 dichroics and emission filters are housed in push & click filter cubes
which are exchanged manually.
The high-NA C-APOCHROMAT objectives specially developed for the LSM technique reach the physical
limit in resolving power, and can be used throughout the 380...900 nm spectral range with the same
high quality, producing brilliant images. Note, for fluorescence correlation spectroscopy the
C-APOCHROMAT objectives are highly recommended.
A two-mirror scanner system, controlled by real time electronics, offers several advantages. The large
deflection angle of the scanning mirrors allows a wide area to be scanned. With a 1.25× objective, the
object area scanned is 11 × 11 mm². The scanning field size can be freely selected between 4 × 1 and
8192 × 8192 pixels.
It is possible to rotate the XY scanning field through 360° and carry out XY scans without having to
rotate the specimen itself under laser radiation load.
For FCS operation all detectors able to count photons can be used for FCS. These are GaAsP PMTs of the
array detector, BiG.2 and the cooled multialkali PMT in the red spectral range. Multialkali PMTs of the
three channel configuration without GaAsP PMTs or NDDs cannot be used for FCS.
Selection of the specimen detail of interest for zooming is fast and convenient, and the zoomed image is
automatically centered. This saves the job of specimen centration with the microscope stage.
Bi-directional scanning doubles the scanning rate to a maximum of 13 frames/sec (at 512 × 512 pixels).
Two fluorescent dyes can be viewed and documented in a quasi-simultaneous mode when using two
different laser wavelengths for bi-directional scanning. This will prevent cross talk between detection
channels.
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