ZEISS Knowledge Base

System Validation with the Particle Test Standard

To check the calibration-setup of your microscope system for a component cleanliness analysis, you can use a test slide (round glass slide) with a defined number of different objects. For this, ZEN core offers the following two test standards:

Particle Test Standard (CC) (2010) Slide D45
Particle Test Standard (CC with OC) (2010) Slide D45

The two standards are available in Manage Templates. You can acquire images of the test slide and use the standards together with the image analysis to determine if your microscope is configured as needed for the component analysis. To get a good and quick impression of the system status, we recommend to only image one group of objects on the slide.

The following example shows an acquisition of the test slide with two polarization channels and the segmentation result with the detected objects inside the drawn measurement frame:

Test slide with Pol 0 channel

Test slide with Pol 90 channel

Segmentation result with detected objects inside the measurement frame

For accessing the information of your analysis, the table(s), report document(s) and particle documents are saved in Browse Results, and can be opened from there.

The following table shows an example result for Metallic Shiny Particles found on the test slide. The particle counting results are based on a measurement area with a diameter of 14.5 mm.

Best Practice for Particle Test Slide and Standard

Apply the best practice recommendation for the preparation each time you perform a measurement using the particle test standard. The configuration of the job template and the dataset generation are only required once in the beginning to receive a meaningful statistic data set for limit value determination. These are later used for routine measurements of the particle test slide. Point three describes how to measure and to interpret your results.

Apply this best practice recommendation each time you perform a measurement using the particle test standard, see Preparation before Using the Particle Test Standard.
Perform the following configuration steps once in the beginning to receive a meaningful statistic data set for limit value determination, see

For information on how to interpret your results, see Routinely Measuring and Interpreting Results.

Preparation before Using the Particle Test Standard

Before you run the Particle Test Standard to check the calibration, carry out the following preparations.

Moisten the slide carefully with a soaked, soft and lint-free cloth (water/detergent solution). Clean the slide afterwards in fresh water. Gently dry the slide with a second dry, soft and lint-free cloth.
We recommend to insert a plain white CN-Derivate filter membrane on the holder underneath the particle test standard. Place the slide with the particles shimmering yellowish on top of the plain filter membrane in the slide holder and turn the top lid carefully until it is fixed.
Mount the filter holder in the stage insert and set a focus on the sample with the large cross in the center.

You have prepared the calibration check using the test slide. If your Job Template is configured, continue with the check, see Routinely Measuring and Interpreting Results.

Configuring the Component Cleanliness Job Template

Configure the job template so that you can run the test standard on a regular basis.

In the Light Path Editing tool, define an appropriate acquisition setting.
Use the 10x objective magnification for the measurement.
Adjust the exposure time best suited for acquisition of the POL-90 image and bright image POL-0, and use the determined exposure time for further measurement.
Apply the shading correction on a particle free area, e.g. close to the cross in the center.
Define a Tile Region with a diameter of 14.5 mm with the cross in the center.
In the Frame Setup tool, set the parameter for the mode to Inside only.
Use the SW-Autofocus for the measurement. A good starting parameter is to focus on each 25th tile.

Generating Dataset

Run the configured job template ten times and archive the results.
Inspect the particle results in the Size Distribution view.
The particles on the slide are found in the absolute counts of the metallic shiny particles, see also the image of the particle gallery below.
Artifacts like fibers or contaminant particles from the environment are detected in the type classes non-shiny particles or fibers.
Calculate the mean value for each particle size class and determine the limit values accordingly. Consider the absolute count of the metallic shiny particles for the limit value determination. See an example for measurement results and the mean value calculation in the table.
Create a new component cleanliness standard template with the determined limit values per particle size class.
This defines your acceptance criterion for the allowed particle number per size class.
Add this standard template to your job template and save it.

Routinely Measuring and Interpreting Results

Use the prepared and pre-configured component cleanliness job template for routine measurement. For this purpose, run the job template once in a certain time period which best suits your needs, e.g. daily, weekly. Inspect the results size in the distribution view.
The particles on the particle test standard are found in the class of absolute counts of the metallic-shiny particles. Artifacts like fibers or contaminant particles from the environment are detected in the type class non-shiny particles and fibers.
The histogram chart and class table document with an ok/n.o.k display the measurement results. The scatter plot shows a regular pattern, see the second image below.

Particle gallery of a test slide analysis

Scatter plot of a test slide analysis (D 14.5 mm)