Particle Size Protocol

Introduction

The Particle Size protocol provides a simple Particle Size Analysis (PSA) workflow. The measured particles may be classified based on diameter or area values for distribution analysis, in addition to reporting D50, D10 and D90 values per image.

Particle Size Protocol Information: 

  • Inputs: One image or image channel containing the particles of interest.
  • Time: End Point 
  • Data Output: 
    • Total Object Count, Diameter (Mean), D50, D10, D90, Diameter (Standard Deviation), Area (Mean), Area (Medan) and Area (Standard Deviation) per image.
    • Classification Statistics: Class Name, Particle Area (Sum) and Particle Count per image.
    • Optional snapshot image of the analyzed location(s).

Particle Size Protocol Setup: Step1

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 particle size analysis can be found in the File/Open/Demo Images in the Protocols Materials/Particle Size 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 Particle Size protocol located in the Materials collection.
  4. The following steps are based on using the using a pre-configured protocol for 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 channels, if your dataset contains multiple channels, select the channel that contains images of particles from the “Particle” combo box.
  7. 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 dropdown.
  8. You may require a Region of Interest (ROI) to define the area of the image to analyze (to avoid calibration bars on the image or limiting the analysis to a specific area or structure).
    Press the Create ROI button under the Region of Interest section.  Draw the ROI and press the OK button in the interactive dialog box when done.  The ROI may also be deleted by pressing the Double Red X button located next to the Create ROI button.

Note: The ROI will be applied to all images when analyzing images individually or in a batch processing mode.

  1. Turn on Interactive Mode and Press the Next Step button.

Particle Size Protocol Segmentation: Step 2

For more information on segmentation and recipe creation, please refer to the Protocols Help section: Protocol Segmentation.

  1. Segmentation
    1. The particle are automatically segmented using the default or saved protocol parameters.
      The protocol is initially setup to use Smart Segmentation to identify       particles. You can change to threshold-based segmentation if you prefer.
      Press the OK button in the Interaction popup when done adjusting the segmentation.
  1. Adjust the segmentation options as needed to identify particles. Press the Apply button to save the changes to the protocol.
    1. 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 and interactive histogram to set the limits.
    2. Clean Borders: removes cells touching the edges of images.
    3. Fill Holes: when selected, any holes within your objects will be filled.
    4. Split: select either Boundary Shape or the Watershed cell splitting methods to separate joined cells or None.
    5. Classify: select either None, Single or Learning method used to classify objects based on morphological and/or intensity parameters.

Press the Next Step button or press the Back button if the segmentation or other steps in the protocol need further adjustment.

Particle Size Protocol: Results

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