Quick visualization of neurons in brain tissues using an optical clearing technique.
Isotropic
Optical clearing
Patch-clamp recording
ScaleSQ
Visualization
Journal
Anatomical science international
ISSN: 1447-073X
Titre abrégé: Anat Sci Int
Pays: Japan
ID NLM: 101154140
Informations de publication
Date de publication:
Mar 2019
Mar 2019
Historique:
received:
20
08
2018
accepted:
08
12
2018
pubmed:
3
1
2019
medline:
6
4
2019
entrez:
3
1
2019
Statut:
ppublish
Résumé
Neurons are classified into several morphological types according to the locations of their somata and the branching patterns of their axons and dendrites. Recent studies suggest that these morphological features are related to their physiological properties, including firing characteristics, responses to neuromodulators, and wiring patterns. Therefore, rapid morphological identification of electrophysiologically recorded neurons promises to advance our understanding of neuronal circuits. One of the most common anatomical cell identification methods is neuronal reconstruction with biocytin delivered through whole-cell patch-clamp pipettes. However, conventional reconstruction methods usually take longer than 24 h and limit the throughput of electrophysiological experiments. Here, we developed a quick, simple cell reconstruction method by optimizing the tissue clearing protocol ScaleSQ. We found that adding 200 mM NaCl almost entirely prevented tissue swelling without compromising optical clearing ability. This solution, termed IsoScaleSQ, allowed us to increase the transparency of the gray matter of 500-µm-thick slices within 30 min, meaning that the total time required to reconstruct whole-cell recorded neurons was reduced to 1 h. This novel method will improve the efficacy and effectiveness of electrophysiological experiments linked to cell morphology.
Identifiants
pubmed: 30600446
doi: 10.1007/s12565-018-00473-z
pii: 10.1007/s12565-018-00473-z
doi:
Substances chimiques
biocytin
G6D6147J22
Lysine
K3Z4F929H6
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
199-208Subventions
Organisme : Grants-in-Aid for Scientific Research
ID : 18H05525
Organisme : the Human Frontier Science Program
ID : RGP0019/2016
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