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Expansion microscopy-based imaging of nuclear structures in cultured cells.

STAR protocols (2021-07-06)
Antoine Gaudreau-Lapierre, Kirk Mulatz, Jean-Claude Béïque, Laura Trinkle-Mulcahy
ABSTRACT

Expansion microscopy is a sample preparation technique in which fixed and immunostained cells or tissues are embedded in a cross-linked network of swellable polyelectrolyte hydrogel that expands isotropically upon addition of deionized water. We utilize the X10 method for tenfold expansion of U2OS cells with concurrent DNA staining. A custom 3D-printed gel cutter and chambered slides minimize gel drift, facilitating analysis of the components of nuclear structures at nanoscale resolution by conventional microscopy or Airyscan confocal imaging. For complete information on the generation and use of this protocol, please refer to Do et al. (2020).

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Sigma-Aldrich
Triton X-100, Molecular Biology
Sigma-Aldrich
TWEEN® 20, viscous liquid, suitable for cell culture
Sigma-Aldrich
N,N,N′,N′-tetrametil-etilendiammina, BioReagent, Molecular Biology, ≥99% (GC)
Sigma-Aldrich
Paraformaldehyde, powder, 95%
Sigma-Aldrich
Proteinasi K, buffered aqueous glycerol solution, Molecular Biology, ≥800 units/mL
Sigma-Aldrich
Sodium acrylate, 97%
Sigma-Aldrich
bisBenzimide H 33342 trihydrochloride, ≥98% (HPLC and TLC)