Published May 19, 2025 | Version v1
Software Open

AgRP and ATG5 Quantification Macro

  • 1. ROR icon Freie Universität Berlin
  • 2. ROR icon Zuse Institute Berlin

Description

This ImageJ macro was developed to perform automated quantitative analysis of microscopy images for the measurement of AgRP (Agouti-related peptide) and ATG5 (autophagy-related protein 5) signals in hypothalamic ARC (arcuate nucleus) regions. The macro processes multi-channel fluorescence images, extracts specific regions of interest (ROI), and calculates mean and max intensity values for AgRP and ATG5 signals.

Key Features:

  1. Batch Processing: Processes all files with a specified suffix (.lif by default) in a given input directory and its subdirectories.
  2. Channel Selection: Allows user-defined selection of channels for AgRP, ATG5, and nuclei staining.
  3. Automated Segmentation: Uses Gaussian blur and Otsu thresholding to segment nuclei and calculate particle-based measurements.
  4. Data Export: Generates a summary CSV file containing image names, AgRP mean, AgRP max, and ATG5 mean values.
  5. Overlay Generation: Creates PNG overlays showing the segmentation results overlaid on the original channels.

Files

AgRP and ATG5 Quantification Macro.pdf

Files (168.6 kB)

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Additional details

Funding

Deutsche Forschungsgemeinschaft
CRC1449 Dynamic Hydrogels at Biointerfaces 431232613
Deutsche Forschungsgemeinschaft
Core Facility BioSupraMol 213868804

Software

Programming language
ImageJ Macro