4.5 Article

Nanometer-scale Multiplexed Super-Resolution Imaging with an Economic 3D-DNA-PAINT Microscope

期刊

CHEMPHYSCHEM
卷 19, 期 22, 页码 3024-3034

出版社

WILEY-V C H VERLAG GMBH
DOI: 10.1002/cphc.201800630

关键词

DNA; nanotechnology; photophysics; single-molecule microscopy; super-resolution imaging

资金

  1. DFG through the Emmy Noether Program [DFG JU 2957/1-1, SFB 1032]
  2. ERC through an ERC Starting Grant (MolMap) [680241]
  3. ERC Proof of Concept Grant (Resolve) [790594]
  4. Max Planck Society
  5. Max Planck Foundation
  6. Center for Nano-science (CeNS)
  7. DFG through the Graduate School of Quantitative Biosciences Munich (QBM)
  8. International Max Planck Research School for Molecular and Cellular Life Sciences (IMPRS-LS)
  9. European Research Council (ERC) [790594] Funding Source: European Research Council (ERC)

向作者/读者索取更多资源

Optical super-resolution microscopy is rapidly changing the way imaging studies in the biological and biomedical sciences are conducted. Due to the unique capability of achieving molecular contrast using fluorescent labels and sub-diffraction resolution down to a few tens of nanometers, super-resolution is developing as an attractive imaging modality. While the increased spatial resolution has already enabled structural studies at unprecedented molecular detail, the wide-spread use of super-resolution approaches as a standard characterization technique in biological laboratories has thus far been prevented by mainly two issues: (1) Intricate sample preparation and image acquisition and (2) costly and complex instrumentation. We here introduce a combination of the recently developed super-resolution technique DNA-PAINT (DNA points accumulation for imaging in nanoscale topography) with an easy-to-replicate, custom-built 3D single-molecule microscope (termed liteTIRF) that is an order of magnitude more economic in cost compared to most commercial systems. We assay the performance of our system using synthetic two- and three-dimensional DNA origami structures and show the applicability to single- and multiplexed cellular imaging.

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