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Introduction
The Particle Analysis module is an option for NanoRule+T™ SPM analysis software and is used for identifying and analyzing particles. There are several provisions for identifying particles. Once identified, many parameters including area, perimeter, height, and length may be calculated. Several options are available for reporting the results from the particle analysis software option.
A particle is defined as a surface entity which is spatially defined as an individual unit. A critical issue for successful particle counting is to distinguish the particles from the image background or "Segmentation". The method employed for detecting segments is called "Threshold Segmentation". As the name indicates, the key requirement for detecting particles is to set a proper threshold level in the Z scale.
The five steps required for counting grains in an image are:
Step1: Prepare Your Image
Step2: Select Particle Analysis Function
Step3: Set The Threshold
Step4: Search and Count Particles
Step5: Report Particle Analysis Results
Step 1: Prepare Your Image
To prepare for particle analysis, an image must be leveled. Because the method we use for counting is "Threshold Segmentation", a leveled image produces a better particle counting result. The standard NanoRule+T™ software leveling functions are used to create an image for particle analysis.
Step 2: Select Particle Analysis Function
The Particle Analysis function can be selected from menu Analysis->Particle Analysis or with the toolbar button.
When the Particle Analysis dialog box is opened, a highlighted image is displayed. The highlighting is defined by a pre-set threshold level and detection mode. The preset threshold may not be optimized for the image, so you may need to adjust this threshold.
Step 3: Set The Threshold
Particle Analysis Dialog Box: In the Particle Analysis Dialog Box, if the Threshold check box is selected, a defined threshold level is applied to the image.
Threshold Edit Window: The threshold window can be opened by clicking the "Edit" button in the Threshold group. An image histogram is displayed. A red/green line in the histogram window indicates the current threshold level. The color bar in the histogram has two regions with two different palettes. One is the original image palette, and one is a highlighted palette.
• If "detect valley" is checked, the lower data range (lower color bar) is the highlighted range.
• In contrast, if "detect peaks" is checked, the higher range is the highlighted range.
Object counting is performed on this highlighted range. To get the optimal threshold level, a user can drag the red/green line with a mouse. The image will be updated with the new threshold level immediately.
Palette: Click the "Palette" button to change the highlighted palette. Either a palette or single color can be chosen.
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Step 4: Search and Count Particles
TCount Button: After a proper threshold level is defined, Click the "Count" button. All objects are defined, highlighted and counted.
Segment Option: The method for selecting segments may be modified. This is done by clicking the “Option” button in the search group. The “Segment Option” dialog box then appears.
Highlight: An object can be highlighted in the following three different styles:
Separated by: Particles can be separated by one of two methods:
• 4-Connected: only objects that share a corner pixel will be counted separately.
• 8-Connected: objects that are connected in any way will be counted as one.
Boundary (in pixel): Adjusts the boundary of the image to determine the inclusion of partial objects.
Smooth: Provides three different filtering kernels for pre-processing the image.
Sort By: Label of the objects can be sorted in area, perimeter, length, or width. The number is ordered from large to small. The default is a searching order.
Sorting by area. The object will be labeled from largest area to smallest area.
After finishing all changes, click Apply button, the new segment option will affect counted objects.
Show Result: After object counting, Results can by shown by checking this box:
Segment Edit: After counting objects, user is able to edit objects in the image.
Measurement: After counting objects, a few parameters will be measured
After counting objects, a few parameters will be measured.
Option: With this option a user is able to choose his own parameters that are displayed in the Report Dialog Box. When an item is selected on the left side, an illustration is presented on the right side.
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Step 5: Report Particle Analysis Results
When the report button is selected, a Measurement Report is displayed. The report items are based on user’s selection from the Measurement Option. Sorting is available for each parameter by clicking the column header.
Export: Export data is available from the context menu (right mouse click on the report window). Data can be exported in Microsoft™ Excel™ format.
Plot: After counting is finalized, all available plots are enabled. Once an item from the plot window is checked, the plot window is displayed.
Plot Function: Users can generate their own set of Plot Functions. When the Modify button is selected, the Plot Function Dialog Box is shown. In the Plot Function Dialog Box, the user can choose the items from the left side to create a plot.
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