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Camera Selection and Alignment

Camera Selection and Alignment

Selecting the camera
You can choose between a LMD CC7000 camera, one of the digital cameras in the Leica DFC range and UCAPI cameras.

LMD version 7.6.0 only supports UCAPI cameras (DFC7000 T, DFC3000 G and DMC2900) and is not compatible with the LMD CC7000
camera. LMD version 7.6.1 only supports the LMD CC7000 camera, but not the UCAPI cameras. Both versions support the TWAIN driver.

Select the option Select Camera Type in the Camera menu:

Check the default setting. If you change it, it will be necessary to re-start the program.

If you select the LMD CC7000 camera and a firewall is activated on your computer, make sure that the microdissection software is added
to the list of exceptions to admit the communication between the program and the LMD CC7000 camera.

By selecting the option Camera Setup Window of the Camera Menu one of the following dialogs for camera settings is opened.

For LMD CC7000:

In Automated mode the Gain value can be set manually. Additionally a White Balance can be activated and an Offset for the black level
can be defined. When the automatic function is switched off, you can also set the Exposure value manually.
Camera Selection and Alignment

For Leica DFC:

For further information on camera settings, please consult the separate camera manual.

If the camera setup is not automated the software will distinguish between the camera settings for the sample/specimen and collector
inspection starting with LMD software V7.6.0. So individual camera setups for the specimen and the collection device can be defined,
depending if you open the camera menu looking to the specimen or collector.

Aligning the camera

For LMD CC7000 camera:
Make sure that the camera is mounted with the Leica logo pointing to the front, that means towards the eyepieces.

If you find that the edges of the live image do not run parallel to the edges of the image window, you can now fine-adjust the camera.
Proceed as follows:

Switch to scanning magnification 5x, 6.3x or 10x, if necessary, using the appropriate keys of the Smartmove control or the x/y joystick.

Focus the microscope image.

Select a prominent specimen detail and move it to the top right corner of the image window.

Move the stage in x direction so that the specimen detail in the image window is moved to the left.

Check that the distance to the edge of the image window remains the same.
If this is not the case, the camera must be rotated accordingly.

Slacken the clamp for the C-mount, rotate the camera and check the alignment again.

Reclamp the C-mount.

Calibrating the Laser

Calibrating the Laser

Calibration of the laser is necessary for each objective for the conversion of the coordinates of the mouse cursor into the coordinates
of the laser beam. The aim is to ensure that the figure drawn by the mouse on the screen is identical with the shape cut out by the laser.

Calibration of the laser is only necessary if there are deviations between the drawing and the cutting line, for example, after disassembly
and reconnection of the laser if it has to be transported. The laser calibration may be repeated at any time and is performed and stored
for each objective separately.

Calibration is performed as follows:

Focus on an empty area of a membrane slide or use an empty membrane slide.

To put a slide on the stage or change slides, press the Unload key in the toolbar. The stage then moves to a suitable position for slide
change. (See also Changing slide).

Check the setting of the laser beam to ensure adequate cutting quality. To do this, press the toolbar key or select the
Control option in the Laser menu.

Calibrating the Laser

The laser power and point distance (speed) parameters can be altered here. The values are preset at the factory, but depend on the
specimen. The values are adapted to the specimen at Specimen Balance. Start with the Factory Settings. Then change the Power and
Speed parameters. Check the setting by making a cut. Keep adjusting the parameters until you see a clean cut.

Now press the Calibrate key and confirm the following message with Yes.

The laser cuts a cross into the top right of the specimen.

Move the mouse cursor to the center of the cross and confirm by clicking it with the left-hand mouse key.

Proceed in the same way for the three other crosses.

Conclude the calibration procedure by clicking OK in the following dialog.

If one or more of these crosses are no longer visible on the screen, check at first the camera alignment. The crosses should form a rect-
angle lying parallel to the edges of the image window.
Calibrating the Collection Device

When the camera has been set up correctly, change the spacing between the calibration crosses in the Options/Settings/Misc dialog.
This is particularly important if the camera has been changed, causing a different picture area to be displayed on the monitor.
If this is not successful, it is necessary to center the laser.
See Problems and solutions.

The calibration procedure can be canceled at every time with Stop Calibration.

Calibration can also be done by selecting the Calibrate option in the Laser menu.

An automated laser calibration can be performed using the Auto Calibrate button. See Laser menu.
The Auto Calibrate function is designed for mirror slides (#11090615012005 ) only and will not work on sample slides.

Calibrating the Collection Device

To insert the collection device in the microdissection stage press the key
The stage moves to the front and the following dialog is opened:

Calibrating the Collection Device

Choose the collection device at the field Device Type:

PCR Tubes (4-fold holder)
8-fold Strip (8-well holder)
Petri (petri dish) or
Universal Holder

Remove the UV stray light shield.

! Attention!
It is essential to remove the shield as when focusing is now carried out without the sample holder, a focal plane is set, a lower focal
plane is set, which could lead to collisions!

Remove the slide holder.

Turn the scanning objective 5x, 6.3x or 10x into the light path.

Click Move to Reference Point in the Adjust Reference Point (RP) field of the Change Collector Device window.
A calibration hole on the collection device will then be moved into the light path automatically. The hole can be seen in the image win-
dow as a bright circular spot on a dark background.

Center the hole by focusing the image and correcting the position, if necessary, by clicking the arrow keys.

Confirm the correct position with OK.

The dialog is closed and the stage moves back to the previous position.

Press again the Unload key to put the slide holder back in position. Then close the dialog. Focus the image as soon as the stage has
reached its previous position.

Now you can replace the UV shield, making sure it clicks into position.