Structured Illumination - Tirf-Filtered - Zeiss LSM 880 Operating Manual

ZEISS
Fig. 497 Structured illumination – TIRF-
Filtered tool selected
Fig. 498 Structured Illumination – TIRF-
Filtered tool
Fig. 499 Error message for falsely loaded
image dimensions in Input image 2
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CHAPTER 1 - SYSTEM OPERATION
Left Tool Area and Hardware Control Tools
000000-2071-464
5.4.17.3 Structured
Filtered
This function works for SIM images (stack) in
combination with a TIRF image. The lowest frame
(first frame) of the stack, preferentially one taken
close to the coverslip, will be taken and combined
with the TIRF. The SIM image contributes the x,y
enhancement, the TIRF image the z-resolution of
the TIRF.
Select the TIRF-Filtered tab to select the TIRF-
Filtered Processing tool (Fig. 497). Activate the
SIM image you want to process in the image
container and press the Select button for Input
image to load the SIM image. Activate the TIRF
image you want to use as the reference in the
image container and press the Select button for
Input image 2. Then expand the Methods
Parameters tool window to have access to setting
parameters (Fig. 498).
You can toggle between Input image 1 and 2 in
the Preview tab window, which also displays a
preview of the output image.
The Image input description field lists all the
channels in the SIM column, the used lasers of a
channel in the Laser column and the employed
camera coded by a number in the Camera
column.
The Image input 2 description field lists all the
channels in the TIRF column and the used laser of
a channel in the Laser column.
It is important to load the SIM image as
Input image and the TIRF image as Input
image 2 as the software expects it that way.
If in Input image 2 an image stack is
selected, the system gives an error message
as it expects a single plane (Fig. 499).
LSM 880
Illumination – TIRF-
10/2014 V_01