Olympus Spinning Disk - DIC images

Start a new Experiment on the "Experiment manager" tab.

Make sure the microscope is properly aligned and that you have checked your Koehler illumination.

Remove any lid you might have above your sample.

Add your observation method DIC to the experiment.

Do not forget to add an additional DIC_OUT observation method at the end to remove the extra components.

An extra DIC_OUT image will be generated, you can delete or ignore it. For more info please contact the ZMB staff.

You can find also find DIC and DIC_OUT observation methods under My functions - "ZMB DIC".

Double click on the observation method box to activate it.

Check the "Synchronize Shutter" box option if you would like to minimize exposure of your sample to light.

Press Live.

Adjust exposure time accordingly.

Under the "Microscope Control tab" your will find the two DIC Polarizers:

Scroll down until you find IX3 DIC slider.

Here you can adjust the orientation of the polarizer.

Make sure your image is not saturated with the help of the Adjust Display pannel.

The Camera can be run at 8 bit, 12 bit as well as 16 bit (recommended).

Here you can toggle the camera bit-depth between 16, and 8-bit and adjust the pixel clock (camera read out speed).

If you are acquiring at 8 bit the histogram max should be lower than 255, at 12 bit lower than 4095, while if you are using 16 bit the max should be lower than 65535.

For more details refer to the different camera modes here: Olympus Spinning Disk - 2: Multichannel + Z-stack acquisition.

Once you are satisfied with your settings right click on the observation method and select "Get settings".

Repeat the procedure for the remaining observation methods.

Name your experiment.

Press Start.

Visualize your image

If you want to acquire fluorescence + DIC images, make sure you remove DIC components from the light path after the DIC image and before acquiring the next fluorescent image, time point, position etc.

Remove DIC components by adding a DIC_OUT observation method.

An extra DIC out image will be generated, you can delete or ignore it. For more info please contact the ZMB staff.

Use the multicolor option to overlay the two channels.

The 2 channels should have the same bit depth, size and format.