Zeiss LSM 880, Operating Manual

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CHAPTER 1 - SYSTEM OPERATION
ZEISS Center Screen Area / Image Containers - Display and Image Analysis LSM 880
524 000000-2071-464 10/2014 V_01
For three of the terms (G
s
, A and G
d
) settings menus are available. To open a settings menu, press the
corresponding Settings button.
Scanning G
s
(
ξ
,
ψ
)
describes the scanning term of
the model function. Scanning is only active in the
Show all
mode. To open the
Scanning
settings
press the Settings button (Fig. 740).
If the
Diffusion coefficient
check box is ticked,
the diffusion coefficient will be fitted, otherwise
the diffusion time. In case the box is checked, the
2 Photon
check box will be accessible. If ticked
diffusion time scales with 1/8
th
of the diffusion
coefficient, if un-ticked with 1/4
th
. The 1/e
2
radius
of the laser beam can be entered within the
ω
r
in
µm input box by typing or using the arrows. Pixel
dwelling times and line times can be entered in the
Pixel time in
µ
s and Line time in µs input boxes.
These values are useful for simulations when fixed.
They will be overwritten with the correct Meta
data from the image, whenever an image is
loaded. If these Meta data are not available, values
have to be provided. In the
Scanning
information
box the formulas for the Scanning
term G
s
(
ξ
,
ψ
) and the conversion formula between
diffusion time
t
d
and diffusion coefficient D are
displayed as well as a description of the parameter
variables.
Offset d describes any scalar added to the
correlation function. It will be necessary, if the
correlation function does not decay to zero.
Amplitude A describes number of particles taking
into account the form of the point spread function
in relation to the studied structure expressed in the
geometric correction factor
γ
. To open the
Amplitude
settings press the
Settings
button
(Fig. 741). You can select a cylindrical PSF with
γ
=1, a two dimensional Gaussian ( 2DG ) PSF with
γ
=0.5, a three dimensional Gaussian ( 3DG ) PSF
with
γ
=0.35 or a Gaussian-Lorentzian (
GL
) PSF with
γ
=0.076 by pressing the respective button.
Alternatively, if Calibrated is active, you can enter a
calibrated value via the
Calibrated input box.
In the
Amplitude information
box the formulas
for the Amplitude term A is displayed as well as a
description of the parameter variables.
Fig. 740 Settings for the Scanning term
Fig. 741 Settings for the Amplitude term