Protocol for matching protein localization to synapse morphology in primary rat neurons by correlative super-resolution microscopy
STAR Protocols. 2024-09-01; 5(3): 103160
DOI: 10.1016/j.xpro.2024.103160
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Cloâtre T(1), Mondin M(2), Sibarita JB(1), Levet F(3), Thoumine O(4).
Author information:
(1)University Bordeaux, CNRS, Interdisciplinary Institute for Neuroscience,
IINS, UMR 5297, 33000 Bordeaux, France.
(2)University Bordeaux, CNRS, INSERM, Bordeaux Imaging Center, BIC, UAR 3420, US
4, 33000 Bordeaux, France.
(3)University Bordeaux, CNRS, Interdisciplinary Institute for Neuroscience,
IINS, UMR 5297, 33000 Bordeaux, France; University Bordeaux, CNRS, INSERM,
Bordeaux Imaging Center, BIC, UAR 3420, US 4, 33000 Bordeaux, France. Electronic
address: .
(4)University Bordeaux, CNRS, Interdisciplinary Institute for Neuroscience,
IINS, UMR 5297, 33000 Bordeaux, France. Electronic address:
.
Super-resolution imaging provides unprecedented visualization of sub-cellular
structures, but the two main techniques used, single-molecule localization
microscopy (SMLM) and stimulated emission depletion (STED), are not easily
reconciled. We present a protocol to super-impose nanoscale protein distribution
reconstructed with SMLM to sub-cellular morphology obtained in STED. We describe
steps for tracking cells on etched coverslips and registering images from two
different microscopes with 30-nm accuracy. In this protocol, synaptic proteins
are mapped in the dendritic spines of primary neurons. For complete details on
the use and execution of this protocol, please refer to Inavalli et al.1.
Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.
DOI: 10.1016/j.xpro.2024.103160
PMCID: PMC11261141
PMID: 38943646
Conflict of interest statement: Declaration of interests The authors declare no
competing interests.