Correction - Zeiss LSM 880 Operating Manual

ZEISS
Fig. 198 Correction field – appearance for
the WF, SIM and Laser WF imaging
modes
Fig. 199 Auto Z Brightness Correction
Adding a new position with a defined set of channel parameters cannot be done while scanning. The
positions don't need to be listed in a specific order, the program will sort the values automatically in the
background.
Move to moves the actual focus to the highlighted Position in the list without scanning. Starting a
Continuous scan then allows setting or correcting the parameters and storing them again hitting Add.
Highlighted positions can be removed (Remove) or the whole list can be deleted (Remove all).
The interpolation between the Positions in the list can be extrapolated (check the box next to
Extrapolate) to the actual first and last slice of a Z-Stack if those are not part of the range of the
Positions in the list.
When Enable Test is active, the settings of the Position list or the interpolated values calculated for the
respective focus position are applied while scanning at single positions of the stack or while focusing
through the stack using the Focus tool or the focus drive of the microscope. This allows controlling the
settings before starting the actual acquisition of the stack.
A position list can be saved and loaded again for further experiments. Use the Save... and Load...
button accordingly. Make sure the Z reference position of the list matches the reference position of the
actual Z stack. It is highly recommended to set the Z focus to zero (ideally at the first or last slice of the
stack) when defining the stack and the correction positions within the Z stack. For a subsequent
experiment the position zero than has to be defined at the position of the first or last slice respectively.
Then the correction parameters will be applied in the desired way. Setting the focus position to zero does
not change the position values of the list!
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CHAPTER 1 - SYSTEM OPERATION
Left Tool Area and Hardware Control Tools
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5.2.15.5

Correction

Expand the Correction field by pressing the
triangle (Fig. 198).
Correction input box allows you to consider the
different refractive indices between the immersion
medium of the objective (n') and the embedding
medium of the specimen (n), which can be set
between 0.5 and 3 by typing or using the arrow
keys. The number should correspond to the ratio
r=n/n'.
Auto Z Brightness Correction (Fig. 199) allows
setting of different values for the scan parameters
Gain (Master), Digital Offset, Digital Gain and
Laser Attenuation of up to 10 freely selectable
slices within a Z-Stack. During the scan procedure,
the values of these parameters are automatically
linearly interpolated between the neighboring
positions within the
interpolation is selected. This function can be
activated and deactivated with the check box Use
Correction.
Add takes the above mentioned parameters and
correlates them with the current Focus Position
which is then added as a Position in the list (for
example the first slice of a stack).
LSM 880
Refractive Index
The
Spline
Z-Stack unless
10/2014 V_01