Horizontal Parallel Layers Protocol

Introduction

The Horizontal Parallel Layers protocol is designed to measure the layer thickness of horizontally oriented parallel layers of materials. 

Horizontal Layers Protocol Information:

  • Inputs: One image containing the sample of interest. 
  • Time: End Point 
  • Data Output: 
    • Layer Measurements: Thickness (Mean), Thickness (Standard Deviation), Thickness (Max) and Thickness (Min).
    • Optional snapshot image of the analyzed location(s).
    • Pie Chart

Horizontal Parallel Layers Protocol Setup: Step 1

For additional information on setting up protocols, please refer to the Protocols Help section: Protocol Setup.

  1. Load your image dataset. An example of a dataset suitable for horizontal parallel layers analysis can be found in the File/Open/Demo Images in the Protocol Materials/Layer Thickness Parallel folder. The following instructions are based on using the demonstration image.
  1. Select the Count/Size tab.
  2. Press the Protocols button to display the protocol browser.
  3. Load the default Horizontal protocol located in the Materials collection under the Materials Layers Parallel group.
  4. The following steps are based on using a pre-configured protocol with the demo dataset. You may also use your own images and settings as needed.
  5. Select the Reset drop down and press the Load Demo button. The Open Demo Image option should also be checked to load the demo image at the same time.
  6. Press the Sync button to load the active image and select the image or image channel containing the image to define the analysis parameters from the “Setup” combo box.
    The demonstration image contains 3 layers oriented from the top of the image going towards the bottom (calibration bar area).  The 3 layers being analyzed are complete coatings located in the middle of the image and to not touch the top or bottom of the image.
    Set the “Setup, ZnPhos, Zinc Rich and Organic Topcoat” identifiers to the Mono image using the combo boxes.

Note: The protocol is designed to define the top layer first and then work downwards towards the bottom of the images.

  1. By default, the Spatial and Intensity selections are set to “Current” and will use the image spatial and intensity calibrations associated with the image if they exist, otherwise the images will be uncalibrated. 
    If an image(s) is uncalibrated and calibrated values are desired, a calibration may be created using the Calibration group located in the Home or Capture tab before running the protocol.  Once an appropriate calibration(s) have been created, they may be selected using the appropriate calibration combo box.
  2. Turn on Interactive and Press the Next Step button.

Horizontal Parallel Layers Protocol Sampling Lines Setup: Step 2

  1. The protocol is preset to measure horizontal layers using a set of specified measurement lines.  Set Layers Orientation to Preset Horizontal using the combo box.
  2. Adjust the Sampling Frequency using the spin buttons to set the number of measurement lines crossing the layers.
  3. Adjust the Offset (%) using the spin buttons to offset the measurement lines from the image sides as percent of the pattern width as needed for the sample.
  4. If the layers are off a few degrees from horizontal, use the Degree Offset spin buttons to align the measurement lines with the layers.
  5. Show Measurements Pattern is on by default to visualize the measurement lines.
  6. Flip analysis direction is off by default and is used to change the direction of the analysis from top to bottom, to starting at the bottom of the image going towards the top.
  7. Press the OK button in the Interaction dialog box.
  8. Press the Next Step button to continue.

Horizontal Parallel Layers Protocol Top Layer Segmentation: Step 3

  1. Each layer is defined by a Top and Bottom Boundary Type.  They boundary type may consists of the following options, which allows the flexibility to either use segmentation or manual drawing methods to define the top and bottom boundaries of the layer. 
    Select the Polygon Top, Polygon Bottom or Polyline to define the Top and Bottom Boundary Layer Type for the top layer.  For the current demonstration protocol, the Top Boundary Type is set to Polygon Top and the Bottom Boundary Type is set to Polygon Bottom.  The Magic Wand is used to create the polygon around the top ZnPhos layer. 
    1. Top and Bottom Boundary Layer: use the combo box to select how the top and bottom layer will be defined.
    1. Polygon Top: Magic Wand, Manual Polygon drawing, or Polygon Auto-trace is used to define the polygon.  The top of the resulting polygon is used to define the top of the top layer in this protocol.
    2. Polygon Bottom: Magic Wand, Manual Polygon drawing, or Polygon Auto-trace is used to define the polygon.  The bottom of the resulting polygon is used to define the bottom of the top layer in this protocol.
    3. Polyline: a manually drawn polyline is used to define the boundary layer.
  1. Select Magic Wand or Manual to define a polygon boundary or the Manual polyline tool to manually draw the boundaries or boundary based on the how the layer boundary is defined in this step. The protocol uses the Magic Wand to create the polygon around the top layer.
    1. Magic Wand: is the default method used to define a polygon around the layer area.  Click in the top layer area to create the polygon.
    1. Histogram: adjust the sliders to change the segmentation range.
    2. Smoothing: low pass filter used to blur image to help identify layer boundary by magic wand.  Only the LUT is changed and not the image intensity values.
    1. Manual: 
    1. Select: activates the Select tool.
    2. Polyline: activates the Polyline drawing tool.  Use the dropdown to select Auto Trace Polyline.
    3. Polygon: activates the Polygon drawing tool.  Use the dropdown to select Auto Trace Polygon.
    4. Split Tool: activates the manual split tool.  Draw splitting lines on the image and click Apply Split button.  Click the button again to cancel the split operation.
    5. Delete: deletes and the selected segmented area on the image. To delete an object, press the Select button and click on the individual object.  Press the Red X Delete button to remove the object.
  1. Press the OK button in the Interaction dialog box when the top layer is correct,
  2. Set the Minimum Area and Fill Holes options as need to define the core center.
    Min Area: Sets the minimum object size to be included in the analysis.  The Edit Range button is used to set the minimum and maximum object size using the interactive histogram to set the limits.
    Fill Holes: turn on the option to fill holes within the layer.   The option is on by default.
  3. Press the Apply button to save the changes to the protocol if the button is active.
  4. For the Horizontal Layers protocol, the Interactive Step is on by default.  The underlying assumption is layers will need to be manually defined.
  5. Press the Next Step button or press the Back button if the segmentation or other steps in the protocol need further adjustment.

Horizontal Parallel Layers Protocol 2nd Layer Segmentation: Step 4

In Step 4, the 2nd middle Zinc Rich layer is defined using the same method and tools as described in the previous Step 3.  Notice in the settings, a new option is present to define the Top Boundary Type.  In this case, the Previous Layer may be used to define the Top Boundary Type of the next layer since the division between the two layers has already been defined by the top layer. 
The bottom boundary of the middle lay may be defined using a polyline tool, since the boundary between middle or 2nd layer is challenging to automatically trace. 

  1. The Polyline drawing tool is selected by default.  Draw a polyline between the dark 2nd layer and the brighter 3rd layer.  Double click to end the drawing.
  2. Press the OK button in the Interaction dialog box to continue.
    Note: If the polyline is not correct, press the Select button and click on the polyline to activate it.  Press the Delete button to remove it and then draw a new polyline by press the Polyline drawing button.
  3. Press the Next Step button or press the Back button if the segmentation or other steps in the protocol need further adjustment.

Horizontal Parallel Layers Protocol 3rd Layer Segmentation: Step 5

In Step 5, the 3rd or bottom Organic Topcoat layer is defined using the same method and tools as described in the previous Steps 3 and 4.  For the demonstration, the Top Boundary Type is set to Previous Layer as before.  However, the 3rd layer may be defined by either drawing a polyline or using the Magic Wand to create a polygon in the area below the 3rd layer. The area below the 3rd layer is unform and may easily segmented using the magic wand tool.  The Bottom Boundary Type is set to Polygon Top.

  1. The magic wand is active and click in in the bright area below the 3rd/bottom layer.
  2. Press the OK button in the Interaction dialog box to continue.
  3. Press the Next Step button or press the Back button if the segmentation or other steps in the protocol need further adjustment.

Horizontal Parallel Layers Protocol Results

For more information on configuring and showing analysis results or saving protocols, please refer to the Protocols Help section: Configuring Results.

Once the resulting data has been configured and displayed, additional images may be analyzed individually or in a batch mode.