Skip to Content
Merck
  • Multiplexed 3D super-resolution imaging of whole cells using spinning disk confocal microscopy and DNA-PAINT.

Multiplexed 3D super-resolution imaging of whole cells using spinning disk confocal microscopy and DNA-PAINT.

Nature communications (2017-12-14)
Florian Schueder, Juanita Lara-Gutiérrez, Brian J Beliveau, Sinem K Saka, Hiroshi M Sasaki, Johannes B Woehrstein, Maximilian T Strauss, Heinrich Grabmayr, Peng Yin, Ralf Jungmann
ABSTRACT

Single-molecule localization microscopy (SMLM) can visualize biological targets on the nanoscale, but complex hardware is required to perform SMLM in thick samples. Here, we combine 3D DNA points accumulation for imaging in nanoscale topography (DNA-PAINT) with spinning disk confocal (SDC) hardware to overcome this limitation. We assay our achievable resolution with two- and three-dimensional DNA origami structures and demonstrate the general applicability by imaging a large variety of cellular targets including proteins, DNA and RNA deep in cells. We achieve multiplexed 3D super-resolution imaging at sample depths up to ~10 µm with up to 20 nm planar and 80 nm axial resolution, now enabling DNA-based super-resolution microscopy in whole cells using standard instrumentation.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Albumin, biotin labeled bovine, lyophilized powder
Sigma-Aldrich
TWEEN® 20, for molecular biology, viscous liquid
Sigma-Aldrich
Protocatechuate 3,4-Dioxygenase from Pseudomonas sp., lyophilized powder, ≥3 units/mg solid
Sigma-Aldrich
Maleimide-PEG2-succinimidyl ester, ≥95%
Sigma-Aldrich
(±)-6-Hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid, 97%
Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
Triton X-100, for molecular biology