This tutorial is one of many instructional videos
presented by the Olympus Industrial Equipment Group.
This tutorial video will provide you with detailed instructions
to capture 3D images and to perform height measurements
within the Surface View add in module
for the Olympus Stream Image-Analysis software.
Within the "Acquisition" tab, start by adjusting the sample
to the bottom level of focus as shown.
You will need to interface with the Process Manager tool window
and be within the Automatic Processes.
Automatic Processes are available
within Stream Motion or Enterprise.
Make sure Instant EFI - "Extended Focal Imaging" is selected,
and for this demonstration only this one process is selected.
Since the image is at the bottom of focus
you will want to select the "Start" set point.
Always start at the bottom level of focus
so the image does not become inverted in 3D view.
You will now need to drive focus
to the top level of the focus range,
or the highest point of your sample.
This can be done using the up arrow buttons
within the "Process Manager"
or by using the hardware control focus knob on the microscope frame.
Once this point has been reached, you must set the "End" point.
You must verify the "Recommended Step Size" value is set.
If the step size is too far above the recommended step size
your image will come out with blur spots.
You can hit the "Apply" button
to have it default to the recommended value
The software sets this value automatically
based on the function of the objective lens.
Once the step size has been determined
it will display the number of Z slices required
for this magnificaiton and Z range.
Once the settings are complete you simply
press the "Start" button at the top.
The Process will now be automated
and perform all of the image captures required
to generate the all in focus image or EFI.
You can watch the progress of the automation
at the bottom of the image window,
or cancel it at any time.
Now that the process is completed you will see the title bar
of the image change to allow you to view all 34 slices of focus points.
You will want to perform a process
to generate the EFI image with a height map.
To perform this select the "Process" Tab,
then select "Enhancement"
and click on "EFI Processing".
The "EFI Processing" window will open.
You can move the preview window or re-size it.
You will want to make sure that the "Height Map" selection is checkmarked
so the processing will generate the height map
for this stack of images based on our Z values.
I also like to have it generate a new document as an output,
so you can always go back to your original if you need to make changes.
If this is not selected it will overwrite the original
document with the "EFI" image.
Once this is done you can simply click OK to perform the process.
Once the process has been performed by the software
you will see an extended focal image, or EFI.
To create the 3D rendering of this image
you have to select the surface view button
at the top title bar of the image.
The surface view option is the module to produce the 3D image
and is a special add in module to Stream Motion or Enterprise.
Now the 3D generated image will be displayed
with the default texture overlay.
You can simply click on the image
and drag it to rotate or tip.
You can also zoom into the image
with the slider control under "Navigation"
To adjust the surface texture and color options
you must click on the "Surface Color Settings" button
at the top of the "Surface View" window.
Within this window you can select
between the source layer, or texture layer.
The source layer is the raw height data in grey scale.
We can go back to the textured layer.
You can also select the color mode from the dropdown.
Single color provides the height data in a single color overlay,
single color height shaded provides the height data
with grey value shading of low to high,
lookup table provides a color band overlay on the height data,
and color from source layer is another grey value height data.
If you select "lookup table"
you can now see height data in the color ramp.
There are several color ramps to select from
by clicking on the open folder icon.
Let's go back to texture from layer.
Even while in the texture mode,
you can still overlay the color ramp with the texture
by selecting "Blend lookup table with other color modes".
This now shows the height data in color ramps,
but also the texture of the sample image.
We can close this window now.
Now go back to single frame view by clicking
on the button at the top title bar of the image.
We will now take some height measurements in Z of the image.
This has to be done in single frame view, not 3D.
It also makes it more accurate for point placement on the surface.
You will need to select the "Processing" tab.
Within the "Measurement and ROI" tool window
select 3D Line measurement mode
Place your first mark on the image.
Then place the second ending mark.
This now shows the 3D Intensity projection
between the two height points selected.
We can now also select a point for measurement
on the opposite side of the sample.
This second point shows the same height measurement
on both sides of the sample as expected.
In this instructional video, we have covered 3D imaging and measurements
within the Olympus Stream Image-Analysis software.
We hope this video has been beneficial,
and we invite you to contact us directly with any questions.