In the Laboratory or Farm Shop
Mulch fragment cleaning and camera preparation
- Wash the collected, sieved plastic mulch fragments to remove any attached soil or organic matter (e.g., plant roots).
- Place each cleaned fragment on a white paper sheet, cover with another white paper sheet, and press with a weight (e.g., books). This step will flatten the fragments, facilitating image area measurement.
- Once the fragments are fully air-dried and flattened, photograph them with a digital camera – a good quality phone camera or a professional camera (e.g., Nikon D5100 with a DX Nikkor 18-55 mm objective lens).
- Mount the camera on a stand 50 cm above the mulch sample, with the camera lens pointing downward vertically.
- Place a ruler next to the fragments in each image to provide a reference scale for subsequent image analysis.
On the computer
- Name each photo so they have identifying file names. For example, use the naming convention “Location SamplingDate Field Replicate Size”. For example, Figure 2 is named “WA B600 post rep4 5mm”.
- Save the photos in JPEG format.

ImageJ analysis
- Open ImageJ.

- Open ‘Image file’, ensure the scale is set correctly by clicking the straight-line icon on the top toolbar, then clicking and dragging to draw a line along the edge of the ruler. You may need to zoom in (Image → Zoom → In) to ensure the line is drawn precisely along a known distance on the ruler (i.e., 10 cm). Click ‘Analyze → Set Scale’. For ‘Known Distance’, enter the length of the line drawn (e.g., 10), and for ‘Unit of Length’, enter cm. Ensure that ‘Global’ is checked, then click ‘OK’.

- Click ‘Image → Adjust → Color Threshold’. This step is used to separate plastic mulch fragments in an image based on their color. Then a new window with a red background will open that allows you to adjust the colors.


- Click ‘Process → Binary → Make Binary’. The thresholded images are then converted to binary format to generate masks for quantitative analysis.

- Draw out the specific image range to be analyzed by clicking the ‘Freehand selections icon’ on the top toolbar. This step is to ensure that the follow-up analysis only focuses on plastic film fragments rather than the ruler and items around the paper.

- Click ‘Edit → Invert’. The binary images are then inverted to ensure that the regions of interest were represented as white objects on a black background prior to quantitative analysis.

- Click ‘Process → Binary → Erode’, to remove edge noise and separate closely adjacent objects prior to quantitative analysis.

- Click ‘Analyze → Set Measurements’, and then check the indices that you want to measure, for example, as shown below. Then, click ‘OK’.

- Click ‘Analyze → Analyze Particles’, show the ‘Outlines’. Then, click ‘OK’.

- A summary box with the measurements will appear. Do not close the summary box. All your measurements will show up in this box and you can conveniently download as an Excel file under ‘File’ → ‘Save as’.



