2025
Lotte J. Herstel Sam de Kater, Corette J. Wieringa
Monitoring Changes in Intracellular Chloride Levels Using the FRET-Based SuperClomeleon Sensor in Organotypic Hippocampal Slices Journal Article
In: Bio-protocol, vol. 15, iss. 5, no. 5, 2025.
Abstract | Links | BibTeX | Tags:
@article{nokey,
title = {Monitoring Changes in Intracellular Chloride Levels Using the FRET-Based SuperClomeleon Sensor in Organotypic Hippocampal Slices},
author = {Sam de Kater, Lotte J. Herstel, Corette J. Wieringa},
url = {https://pmc.ncbi.nlm.nih.gov/articles/PMC11896781/},
doi = {10.21769/BioProtoc.5229},
year = {2025},
date = {2025-03-05},
urldate = {2025-03-05},
journal = {Bio-protocol},
volume = {15},
number = {5},
issue = {5},
abstract = {The reduction in intracellular neuronal chloride concentration is a crucial event during neurodevelopment that
shifts GABAergic signaling from depolarizing to hyperpolarizing. Alterations in chloride homeostasis are
implicated in numerous neurodevelopmental disorders, including autism spectrum disorder (ASD). Recent
advancements in biosensor technology allow the simultaneous determination of intracellular chloride
concentration of multiple neurons. Here, we describe an optimized protocol for the use of the ratiometric
chloride sensor SuperClomeleon (SClm) in organotypic hippocampal slices. We record chloride levels as
fluorescence responses of the SClm sensor using two-photon microscopy. We discuss how the SClm sensor can
be effectively delivered to specific cell types using virus-mediated transduction and describe the calibration
procedure to determine the chloride concentration from SClm sensor responses.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The reduction in intracellular neuronal chloride concentration is a crucial event during neurodevelopment that
shifts GABAergic signaling from depolarizing to hyperpolarizing. Alterations in chloride homeostasis are
implicated in numerous neurodevelopmental disorders, including autism spectrum disorder (ASD). Recent
advancements in biosensor technology allow the simultaneous determination of intracellular chloride
concentration of multiple neurons. Here, we describe an optimized protocol for the use of the ratiometric
chloride sensor SuperClomeleon (SClm) in organotypic hippocampal slices. We record chloride levels as
fluorescence responses of the SClm sensor using two-photon microscopy. We discuss how the SClm sensor can
be effectively delivered to specific cell types using virus-mediated transduction and describe the calibration
procedure to determine the chloride concentration from SClm sensor responses.
shifts GABAergic signaling from depolarizing to hyperpolarizing. Alterations in chloride homeostasis are
implicated in numerous neurodevelopmental disorders, including autism spectrum disorder (ASD). Recent
advancements in biosensor technology allow the simultaneous determination of intracellular chloride
concentration of multiple neurons. Here, we describe an optimized protocol for the use of the ratiometric
chloride sensor SuperClomeleon (SClm) in organotypic hippocampal slices. We record chloride levels as
fluorescence responses of the SClm sensor using two-photon microscopy. We discuss how the SClm sensor can
be effectively delivered to specific cell types using virus-mediated transduction and describe the calibration
procedure to determine the chloride concentration from SClm sensor responses.