Dear confocalists,

there has been an interesting discussion during the last days and I would like to answer a few questions arisen on the board for the LSM 510,
 especially concerning Multitracking and spectral option.

Multitracking is an automatic sequentiall measurement of up to 4 so called individual tracks. Every track represents a configuration of the
system covering settings for detection channels, detection beam path, laser lines, pinhole parameters etc., in the end all settings you can
 use to configure the system. The switching between the tracks can be done on a linewise or framewise basis, to speed up it works also bidirectional.
That means as a standard example: On the forward scanner way 488nm only is active with a FITC detection channel (longpass filter), on the reverse
 way over the sample 543nm is active with Rhodamin detection (again with a long pass). Due to the selectivity for the different dye labels on the
excitation site there is no cross talk and you can detect the full emission range of every dyes without need to narrow the spectral detection bandwidth.
 For the bidirectional use there is an automatic offset calibration for phase correction in x and y to secure perfect overlapping of the scan directions.
Some application has been described by Roland Nitschke, the major advantage is of course to exclude bleed through, for instance to seperate
CFP and YFP. Within the 4 tracks you can capture up to 8 channels (4 of them simultaneously in a single track), in case you include online ratio
calculation you can measure the input channels for the ratios in addition.
The Multitracking works also with the new LSM 5 PASCAL. Interesting point is that you can use individual pinhole setting in every track
(motorized and software controlled) and even with a single detection channel one can get multi color images sequentially in an automac way.
The definition and all setting for a multitrack can be done and optimized online during image acquisition for all track which makes it really simple.
 A ones defined multitrack can be stored as a so called "recording" and completely reactivated with a single mouse click.

For the spectral option a confocal channel  can be coupled into a spectrometer via optical fibre (for Chip Montrose' comment: it is really confocal
with a pinhole). This fibre out for a confocal detection channel is a standard interface which could also be used for other detection modes.
The spectrometer allows to get a full spectrum with a single shot for every part of the sample ranging from a single pixel over single or multiple
free hand drawn ROIs or a full frame. There is a range of different spectrometer types depending on the applicational needs from UV to IR.
The spectrometer can be used for FRET measurements and every other spectral measurement.

Best regards
Ronald


Dr. Ronald Wendenburg
Product Manager Laser Scanning Microscopy
Carl Zeiss Jena GmbH, Micro Detection Systems
phone: 0049 (0)3641 64-2646   fax: -3144
e-mail: [log in to unmask]