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Updated: Oct 25, 2025
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Monitoring Neuronal Survival via Longitudinal Fluorescence Microscopy
Published on: January 19, 2019
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A Scalable Method to Study Neuronal Survival in Primary Neuronal Culture with Single-cell and Real-Time Resolution
Ángela Rodríguez-Prieto1, Ana González-Manteiga1, Yaiza Domínguez-Canterla1
1Laboratory of Cortical Circuits in Health and Disease, Centro de Investigación Príncipe Felipe.
Journal of Visualized Experiments : Jove
|August 9, 2021
Summary
This study introduces an affordable and effective method to analyze neuronal survival under toxic stress using sparse fluorescent labeling and automated live imaging. This approach aids in understanding neurodegenerative diseases and facilitates high-throughput drug screening.
Area of Science:
- Neuroscience
- Cell Biology
- Biotechnology
Background:
- Neuronal loss is central to neuropathologies like stroke, Alzheimer's, and Parkinson's diseases.
- Existing methods for studying neuronal survival lack single-cell resolution or are complex and expensive.
- There is a need for accessible and effective in vitro models to study neurotoxicity.
Purpose of the Study:
- To develop a versatile, inexpensive, and effective experimental paradigm for studying neuronal survival under cytotoxic stress.
- To enable automated quantification of neuronal survival using live imaging and sparse fluorescent labeling.
- To provide an adaptable platform for investigating specific molecular pathways and for high-throughput drug screening.
Main Methods:
- Sparse fluorescent labeling of neurons via electroporation, co-cultured with non-electroporated neurons.
- Live imaging and automated quantification of labeled neurons.
- Utilized an oxygen-glucose deprivation (OGD) assay in a homemade hypoxic chamber for a stroke model.
- Image analysis performed using IN Cell Analyzer 2200 or open-source ImageJ.
Main Results:
- Demonstrated a simple, robust, and automated quantification of neuronal survival through sparse labeling.
- Successfully implemented an affordable in vitro neurotoxicity model (OGD assay).
- Developed adaptable workflows for semi-automatic data processing suitable for various toxicity models.
Conclusions:
- The described protocol offers an approachable, affordable, and effective in vitro model for neurotoxicity studies.
- This method is suitable for testing the roles of specific genes and pathways in neuronal survival.
- The protocol can be scaled for high-throughput drug screening in neurodegenerative disease research.
