Expansion microscopy-based imaging of nuclear structures in cultured cells

• Published in: STAR protocols Vol. 2; no. 3; p. 100630
• Main Authors: Gaudreau-Lapierre, Antoine, Mulatz, Kirk, Béïque, Jean-Claude, Trinkle-Mulcahy, Laura
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 17.09.2021; Elsevier
• Subjects: Cell Biology; Microscopy; Protocol; Microscopy; Cell Biology
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2021.100630

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). [Display omitted] •Tenfold expansion of fixed and stained cultured cells with concurrent DNA staining•Custom 3D-printed gel cutter to minimize drift in chambered slide during imaging•Airyscan and Imaris-based imaging and volume rendering of nuclear structures•Approach to calculate expansion factor using Fiji open-source software 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.