MULTIPHOTON LASER SCANNING MICROSCOPY
LSM 510 META NLO
Using the LSM 510 NLO direct coupled system
Carl Zeiss
03/06
B 45-0021 e
9-13
9.3
Using the LSM 510 NLO direct coupled system:
Practical considerations for optimal imaging
9.3.1
Coupling an ultrafast laser to the LSM 510 NLO
Coupling an ultrafast laser to a laser scanning microscope is a fairly simple matter, if several key
considerations are understood. Typically, the routing kit for coupling the LSM 510 NLO to the laser will
consist of the following components: an Acousto-Optic Modulator (AOM), used to control the intensity
of the beam and for beam blanking (allowing for region of interest (ROI) and bi-directional scanning and
for performing Auto-Z-Brightness correction); two 45º mirrors for directing the beam to the microscope
on the table; and a 1 meter lens. In the case of an upright system, the beam is brought up into the scan
head via a periscope, whereas for an inverted microscope, the height of the beam will be adjusted by a
smaller periscope positioned in place of the second turning mirror. These devices ensure that the beam
remains parallel to the table when horizontal and enters the scan head at the proper height (see
diagram). The 45º routing or turning mirrors are adjustable and will be used routinely to peak the
coupling of the beam into the scan head. The 1 m lens is used to expand the beam to fill the back
aperture of the objective. It is mounted in a fixed position by a trained service engineer during installation
and should not be adjusted by the user (Fig. 9-4).
Fig. 9-4
Typical alignment scheme for the LSM 510 NLO direct-coupled system
using and upright(left) or inverted(right) microscope.
