HPLC - ImageJ Protocol

Author

Anna Schweiger

Updated

31 January 2022 by Cathleen Lapadat

Goal

This protocol details the steps to process images of leaf samples measured for HPLC analysis and applicable for SLA.

Supplies/Equipment Needed

• ImageJ software

ImageJ

1. Open image: file - open

2. Set scale:

   1. Click line:
      1. Draw line from one side of the image to the other
   2. Analyze - set scale:
      1. For inner line of boat 0.0028m, set to global, okay

3. Identify leaf

   1. Image - adjust - color threshold
      1. Unselect black background
      2. play with hue, saturation, brightness: leaf should be red
   2. close color threshold

4. Calculate area of leaf

   1. Analyze - analyze particles:
      1. Set lower size limit - Infinity
      2. Show: outlines
      3. Display results
      4. Exclude on edges
      5. Include holes
      6. Summarize
5. Add area to excel
   1. Make sure outlines are of leaf/needle.

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Last update: December 29, 2022