This tutorial is one of many instructional videos
presented by the Olympus America Scientific Equipment Group
Industrial Microscopes
This video will provide you with detailed instructions
on how to utilize the macro manager tool
in the Olympus Stream image analysis software platform
First, we need to create a tool window for the Macro Manager.
This can be easily accessed from the menu View,
Tool Windows, Macro Manager.
You will now notice a tool window
on your desktop for the Macro Manager.
First, we're going to go ahead and click the button "Create Macro."
Type in a name for your macro.
We'll call this one "Particle Detection."
After you have entered a name for your macro
the macro recorder will automatically start recording
every key stroke you make.
You will also notice that the record button is depressed by default
in the Macro Manager tool bar, and is grayed out.
The first step of this macro is to binarize the image.
In other words, convert each pixel
into either black or white.
An image can be binarized from the menu Image, Mode, Binary.
Now, we can select which gray scale values are binarized
to either black or white via gray scale thresholding.
After you have selected which gray scale values
you wish to binarize accordingly,
simply press the "OK" button.
After the binarization has been completed,
you may notice that some neighboring particles may be touching.
Unfortunately, these particles may be inaccurately counted
as one during the particle detection.
Therefore, a morphological filter may be applied,
which will separate these touching particles, automatically.
You can execute this filter from the menu Process,
Morphological Filter, Separate Objects.
Modify the parameters of your particle separation filter,
then click the "OK" button.
Now that the particles which have been previously touching
have been automatically separated,
we can continue onto the next step of the particle detection,
which is to set our gray scale thresholds.
Setting thresholds is the process in which we segment
the image into two distinct components.
One of which are the particles to be detected,
and the other is simply the background,
which will not be detected.
We can automatically segment our image
by selecting our background, whether it's dark, bright,
or have the software automatically detect the background.
After we segment our image, or set our thresholds,
we can simply press the "Count and Measure" button.
After clicking the "Count and Measure" button,
measurement results are listed for each individual particle,
in compliance with the parameters that were previously defined.
For example, area, diameter, or aspect ratio.
Measurement results can then be exported
to Microsoft Excel with the click of a button.
Simply define a file path and file name,
then click the "Save" button.
The workflow is now complete,
therefore you can stop recording the macro
by pressing the "Stop Macro Recording" button.
To run this macro, or any pre-recorded macro,
select it from the drop-down menu, then press play.
The macro will now execute, stepping through
each of the commands listed for that particular macro.
As your application requirements may change,
the macro can be easily modified.
For example, let's say that you no longer wish
to have objects separated in the macro.
Simply uncheck the "Active" box for that particular step.
Next time the marco is ran, that step will be skipped.
Also, the parameters of any step of the macro can be modified.
For example, let's say that the gray scale threshold parameters
of the segmentation are too aggressive and need to be decreased.
By clicking the parameters button,
you can modify the grayscale thresholds accordingly.
Next time the macro is ran, these newly defined parameters
will be utilized, replacing the former.
However, in some instances, there may be no universal parameter
that will work for all images.
Therefore, you may choose to enable the execution mode.
With the execution mode enabled for a particular step,
the macro will pause, prompting the operator
to interactively define the parameters for that particular image on the fly.
When the macro is ran, you will now notice
that the operator is prompted
to set the segmentation gray scale thresholds.
By default, the macro is only available to the author.
However, the author can choose to publish their macro
so that other users, logged on to the same computer,
can access the macro.
Click on the macro management drop down menu.
Then, select "Publish Macros."
Now, check the box for the particular macro
or macros that you wish to publish, the click "OK."
The icon of the double head on the left of the macro name
is an indication that the macro has been published.
In addition to publishing macros on a single computer,
you can choose to export the macro file to a network location,
so that Stream users on other computers can import the macro file.
Further, the user can preview the effects
of each individual step of a macro at their own pace
by clicking the "Step Through Macro" command.
You can create a new macro by clicking the "New Macro" button,
or delete the selected macro by clicking the "Delete Macro" button.
You can also toggle between macros
from the drop-down menu next to the macro name,
where a complete list of all available macros will appear.
In this instructional video, we have covered
how to utilize the Macro Manager tool
in the Olympus Stream Image-analysis software platform.
We hope this video has been beneficial,
and we invite you to contact us directly with any questions.