Skip to Content
Merck
  • Super-resolution dynamic imaging of dendritic spines using a low-affinity photoconvertible actin probe.

Super-resolution dynamic imaging of dendritic spines using a low-affinity photoconvertible actin probe.

PloS one (2011-01-26)
Ignacio Izeddin, Christian G Specht, Mickaël Lelek, Xavier Darzacq, Antoine Triller, Christophe Zimmer, Maxime Dahan
ABSTRACT

The actin cytoskeleton of dendritic spines plays a key role in morphological aspects of synaptic plasticity. The detailed analysis of the spine structure and dynamics in live neurons, however, has been hampered by the diffraction-limited resolution of conventional fluorescence microscopy. The advent of nanoscopic imaging techniques thus holds great promise for the study of these processes. We implemented a strategy for the visualization of morphological changes of dendritic spines over tens of minutes at a lateral resolution of 25 to 65 nm. We have generated a low-affinity photoconvertible probe, capable of reversibly binding to actin and thus allowing long-term photoactivated localization microscopy of the spine cytoskeleton. Using this approach, we resolve structural parameters of spines and record their long-term dynamics at a temporal resolution below one minute. Furthermore, we have determined changes in the spine morphology in response to pharmacologically induced synaptic activity and quantified the actin redistribution underlying these changes. By combining PALM imaging with quantum dot tracking, we could also simultaneously visualize the cytoskeleton and the spine membrane, allowing us to record complementary information on the morphological changes of the spines at super-resolution.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, essentially γ-globulin free, ≥98% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, ≥96% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, Cohn Fraction V, lyophilized powder, ≥96% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, ≥98% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, Protease, essentially free, ≥98% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, essentially globulin free, low endotoxin, ≥98% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, essentially fatty acid free, ≥96% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, protease free, pH 7, ≥98%
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, essentially fatty acid free and essentially globulin free, ≥99% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, powder, BioXtra
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, lyophilized powder, essentially fatty acid free, ≥98% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, low endotoxin, pH 7, ≥98%
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, 1× crystallized, ≥97% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, essentially globulin free, ≥99% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, protease free, suitable for hybridization, pH 7, ≥98%
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, suitable for RIA, pH 5.2, ≥96%
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, low endotoxin, BioReagent, suitable for cell culture, ≥98% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, protease free, essentially globulin free, pH 7, ≥98%
Sigma-Aldrich
Bovine Serum Albumin, For use as a marker in SDS-PAGE
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, pH 5.2, ≥96%
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, pH 5.2, ≥96%
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, pH 7, ≥98%
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, microbiologically tested, ≥96% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, dimer mol wt ~132 kDa
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, essentially IgG free, ≥96% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, fatty acid free, low endotoxin, lyophilized powder, BioReagent, suitable for cell culture, ≥96% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, pH 7, ≥98%
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, ≥96% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, cold ethanol fraction, pH 5.2, ≥96%