Zeiss LSM 880 Operating Manual Download

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CHAPTER 1 - SYSTEM OPERATION

LSM 880

Left Tool Area and Hardware Control Tools

ZEISS

10/2014 V_01

000000-2071-464

377

View 1

is the master view and cannot be moved during the process, the button

View 1

is therefore

inactive and grayed out. Any of the other views can be activated by pressing the according button (e.g.

View2

, Fig. 528). The activated view is automatically displayed in the image (the associated channel is

turend on). To move the view relatively to View 1 the sliders and spin boxes can be used;

for the x-axis,

for the y-axis,

for the z-axis and

Angle

for a roation. The axis that is not present in the maximum

intensity projection is grayed out and inactive (

for the

Front View

and

for the

Side View

Addintionally to the rotation

Angle

, the position of the rotation point can be defined. This is done by

positioning the

red Cross overlay

within the maximum intensity projection image window to the desired

position, the view should be rotated around (Fig. 530).
The

Reset

button moves all views back to the position they were at, as the maximum intensity projection

was generated.
Each view should be activated once with its corresponding button and positioned, resulting in a matching
overlay to

View 1

. Afterwards the maximum intensity projection is switched to either

Front View

Side View

; displayed in the same image window.

Again all views must be moved using the according sliders

and

Angle

to produce a perfect overlay

to View 1.
Switching from

Front View

Side View

should be repeated at least twice, to adust the positioning of

the views.
To controll if the manual alignment is satisfactory, the

Preview

button produces a new image of the

channel and the time point, which is registered with the provided settings. The

Preview

image will

contain fused views, when

Mean Fusion

has been selected at the

Fusion Method

drop down list.

If the result is satisfactory, one can proceed with the remaining settings for

Multiview Processing

If the data was acquired with dual side illumination, which has not yet been fused, the method to fuse
both illumination sides is available in the

Dual Side Fusion

drop-down menu (

Mean Fusion

DFT

Fusion

Maximum Fusion

Mean Fusion

: The average intensity for each pixel is calculated using the values of both illumination

sides.

DFT Fusion

(Discrete Fourier Transform Fusion): The Fusion is performed based upon the published

method ("Multiple imaging axis microscopy improves resolution for thick-sample applications", Jim
Swoger, Jan Huisken, and Ernst H. K. Stelzer, OPTICS LETTERS / Vol. 28, No. 18 / September 15, 2003).

Fine structures and details are more visible after the fusion, which might be worsened when using Mean
Fusion. However, this process can create noticeable artifacts in the resulting image.

Maximum Fusion:

The intensity of each pixel of both illumination images is compared and the value

with the higher intensity is used for the resulting image.

For

Mean Fusion

and

Maximum Fusion

the checkbox

Fusion Subset in X with Preview

can be

activated (Fig. 531). With these controls it is possible to define which area of the left and the right

illumination image should be used for fusion.
When this function is chosen, the image is displayed in the

Preview Tab

in the image container

(Fig. 532). In this preview mode the left hand image shows the original dataset and the right hand image
the preview of processing. Using the

dimension sliders

within the

image control area

the

Z-Position

Illumination

side and

View

can be chosen.

Fusion Subset in X with Preview

the currently chosen View is displayed in the

Current View

box.

With the

Left

slider or input box the area of the left illumination side is chosen. This is marked at the

slider itself with a red line, the input box shows the pixel value along the y axis. Furthermore the cut off is
shown in the preview image with a red line for easy orientation. It is best to set the

illumination slider

of the

image control area

to 1, showing the left side illumination of the original data.