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Quantitative Analysis of Rat Dorsal Root Ganglion Neurons Cultured on Microelectrode Arrays Based on Fluorescence
João Fernando Mari1,2, José Hiroki Saito2,3, Amanda Ferreira Neves4,5
1* Instituto de Ciências Exatas e Tecnológicas - Universidade Federal de Viçosa, 38810-000 Rio Paranaí, MG, Brazil.
International Journal of Neural Systems
|October 30, 2015
Summary
This study presents a new image analysis framework for neuron cultures on microelectrode arrays (MEAs). It enables quantitative morphological analysis, providing insights into cell health and spatial distribution for better electrophysiological studies.
Area of Science:
- Neuroscience
- Biomedical Engineering
- Image Analysis
Background:
- Microelectrode Arrays (MEAs) are crucial for long-term electrophysiological recordings of in vitro neuron cultures.
- Current analysis of MEA neuron cultures often relies on qualitative assessments of microscopy images.
- A need exists for standardized, quantitative image analysis to complement electrophysiological data.
Purpose of the Study:
- To develop and validate a computational framework for quantitative and morphological analysis of neuron cultures on MEAs using fluorescence and transmitted light microscopy.
- To enable standardized image processing and extract key quantitative measures for integrated signal-image studies.
- To assess the culture's state, including cell count, morphology, and spatial relationships with microelectrodes.
Main Methods:
- Neuron segmentation from fluorescence images using thresholding, watershed transform, and object classification.
- Microelectrode localization in transmitted light images via the circular Hough transform.
- Application to dissociated rat dorsal root ganglion (DRG) neuronal cell cultures.
Main Results:
- The framework successfully quantifies population count, neuron-neuron/microelectrode distances, soma morphology, and spatial distributions.
- Achieved a high correct segmentation rate of up to 84% for neurons.
- Demonstrated robust microelectrode identification and neuron segmentation/classification.
Conclusions:
- The developed framework provides essential quantitative data for integrated electrophysiological signal and image analysis.
- It standardizes image processing and offers valuable insights into the physical state of neuron cultures on MEAs.
- The method is adaptable for analyzing cortical and hippocampal neuron cultures with minor parameter adjustments.
