This video demonstrates the use of fluorescence imaging to measure cytosolic calcium levels in short-term and long-term rat hippocampal cultures. By comparing young and aged neurons, it highlights how increased L-type voltage-gated calcium channel density and prolonged activation in aged neurons result in calcium overloading, a hallmark of neuronal aging. Fluorescent calcium indicators reveal these differences through changes in fluorescence intensity upon stimulation.
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Method Article
In Vitro Calcium Imaging in Short-Term and Long-Term Rat Hippocampal Cultures
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May 29th, 2025
In This Article
Abstract
Protocol
1. Short and Long-term Culture of Rat Hippocampal Neurons
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Results
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Materials
| Name | Company | Catalog Number | Comments |
|---|---|---|---|
| Neurobasal Culture Medium | |||
| HBSS medium | |||
| Ham's F-12 medium | |||
| DNase I (from bovine pancreas) | |||
| Fetal Bobine Serum | |||
| B27 | |||
| Gentamicin | |||
| L-glutamine | |||
| 12 mm glass coverslips | |||
| Papain | |||
| 4-well multidish plaques | |||
| Petri dishes | |||
| Sterile pipettes | |||
| Fura2-AM | |||
| Lipofectamine2000 | |||
| NMDA | |||
| Glycine | |||
| Zeiss Axiovert S100 TV microscope | |||
| Xcite ilumination system | |||
| ORCA ER fluorescence camera | |||
| VC-8 valve controller | |||
| SH-27B heating system | |||
| Aquacosmos Software |
