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Updated: Aug 14, 2026
Analysis of Immune Cells in Single Sciatic Nerves and Dorsal Root Ganglion from a Single Mouse Using Flow Cytometry
Published on: December 6, 2017
Open-source method of image cytometry in dorsal root ganglia tissue with immunofluorescence
Michael B Anderson1, J Thomas Curtis1, Kenneth E Miller1
1Michael Anderson, Oklahoma State University Center for Health Sciences, Anatomy & Cell Biology, (E-453/461), 1111 W 17th St, Tulsa, OK, 74135, USA.
Insights
This study introduces a novel automated image analysis method for dorsal root ganglion (DRG) neurons, improving efficiency and standardization in neuroinflammatory pain research. The Image Cytometry in DRG (IC-DRG) approach overcomes manual limitations for accurate protein and antigen evaluation.
Area of Science:
- Neuroscience
- Immunology
- Biotechnology
Background:
- Immunohistochemistry (IHC) is crucial for analyzing proteins in tissues for research.
- Studying dorsal root ganglion (DRG) neurons is key to understanding neuroinflammation and pain signaling.
- Manual analysis of IHC in DRG neurons is time-consuming and prone to errors.
Purpose of the Study:
- To develop a semi-automated, standardized method for analyzing IHC in DRG neurons.
- To overcome the limitations of manual tracing in neuroinflammatory pain research.
- To introduce Image Cytometry in rat DRG (IC-DRG) for efficient neuronal analysis.
Main Methods:
- Utilized a neuronal cytoplasmic reporter specific to DRG neuronal soma.
- Developed a semi-automated algorithm-based approach called Image Cytometry in rat DRG (IC-DRG).
- Employed an ImageJ/FIJI script for automated image analysis, generating nuclear and somatic masks.
Main Results:
- The IC-DRG approach provides binary masks for DRG neuron boundaries.
- Automated analysis successfully overcomes morphological variability and imaging artifacts.
- The method demonstrates high accuracy (94%) when compared to manual scoring.
Conclusions:
- Automated image analysis using IC-DRG offers a significant improvement over manual methods for DRG neuron studies.
- This novel approach enhances standardization and efficiency in neuroinflammatory pain research.
- The ImageJ/FIJI script provides a robust tool for analyzing immunofluorescence in DRG neurons.
Abstract:
Immunohistochemistry (IHC) is a valuable tool in clinical and biological research for evaluating proteins and other antigens in spatially bound tissue. In neuroinflammatory pain research, primary afferent neurons of the dorsal root ganglion (DRG) are studied to understand molecular signaling mechanisms involved in nociception (pain) and inflammation. Measuring IHC (immunofluorescence) in DRG neurons requires manual hand tracing of nuclear and somatic boundaries, which is laborious, error-prone, and may require several weeks to collect the appropriate sample size with a mouse or pen-input display monitor. To overcome these limitations and increase standardization of sampling and measurement, we employed a reliable neuronal cytoplasmic reporter, exclusive to DRG neuronal soma, in a semi-automated algorithm-based approach of Image Cytometry in rat DRG (IC-DRG). The resulting output images are binary nuclear and somatic masks of DRG neurons, defining boundaries of measurement for CellProfiler and manually scored at 94% accurate. Herein, we successfully show a novel approach of automated image analysis for DRG neurons using a robust ImageJ/FIJI script, overcoming morphological variability and imaging artifacts native to imaging frozen tissue sections processed with immunofluorescence.
