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

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.

SUBMITTER: Schueder F

PROVIDER: S-EPMC5727263 | biostudies-literature | 2017 Dec

REPOSITORIES: biostudies-literature

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

Schueder Florian F Lara-Gutiérrez Juanita J Beliveau Brian J BJ Saka Sinem K SK Sasaki Hiroshi M HM Woehrstein Johannes B JB Strauss Maximilian T MT Grabmayr Heinrich H Yin Peng P Jungmann Ralf R

Nature communications 20171212 1

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 ...[more]

PMID: 29233999

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