6-2 Operation
DeepSee Component
compensation provided for each wavelength is governed by the position of the DeepSee
motor that moves optical material on a stage within the beam path.
The motor position, as displayed on the GUI screen, is measured in units of percent of
full motor travel. At each wavelength there is only a certain range of motor positions
available that are determined by upper and lower “soft limits.” The accessible motor
position range at each wavelength is between about 25 to 30 percent, again depending
on the wavelength. These motor limits prevent the beam from missing the optical
elements in the DeepSee component in order to maintain beam transmission through the
DeepSee component no matter how far the motor is commanded to move.
The laser stores calibration files, called objective tables, that correlate wavelength to
motor position. Typically, the operator selects the objective table that best matches their
equipment, and that table is displayed on the controller screen. When the InSight
X3
wavelength is changed, the motor moves according to the currently active table. These
tables usually store just a few calibration points, and the motor positions for
wavelengths between these calibration points are calculated by linear interpolation
using the two closest points. The operator can create, modify, calibrate, and delete
objective tables.
The operator can change the InSight X3 wavelength via the GUI control software or a
user-written program.
Objective Tables
A DeepSee objective calibration file comprises a microscope magnification factor, an
immersion selection (water, glycol, air, or oil) and a numeric aperture. This “lens” is
further defined by including a table containing several points on a graph where the
wavelength at each point is matched to the motor position that maximizes fluorescence
at the target.
When the InSight X3 wavelength is changed, the DeepSee component adjusts according
to the objective table selected.
Creating and Using an Objective Table
For an experiment using a particular optical setup, the operator can select an objective
table from a list of stored objectives or create a new one with new calibration data. The
operator can create a new objective table by cloning it from an existing one, then
modifying it accordingly. Except for the read-only template (a special objective used as
a starting point), the operator can create each objective table, then modify, save, erase,
or replace it. A total of 200 objective tables can be created.
An objective table must contain at least three calibration points, but it is usually
calibrated at several wavelengths in the range of interest. The maximum number of
points is limited to the number of integer wavelengths in the range of the InSight
X3
laser. For example, the objective table for a wavelength range of 680 to 1300
nm, is 620
points (one for each 1
nm change). The operator calibrates by creating a new objective
and adjusting the motor position (or point) to optimize the fluorescence signal at the
target at each wavelength of interest, and then saving these points in the table. Finally,
the operator saves the table as a new objective. Usually 30
– 50 nm increments between
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