124-Color Super-resolution Imaging by Engineering DNA-PAINT Blinking Kinetics.
Orsolya K WadeJohannes B WoehrsteinPhilipp C NickelsSebastian StraussFlorian StehrJohannes SteinFlorian SchuederMaximilian T StraussMahipal GanjiJoerg SchnitzbauerHeinrich GrabmayrPeng YinPetra SchwilleRalf JungmannPublished in: Nano letters (2019)
Optical super-resolution techniques reach unprecedented spatial resolution down to a few nanometers. However, efficient multiplexing strategies for the simultaneous detection of hundreds of molecular species are still elusive. Here, we introduce an entirely new approach to multiplexed super-resolution microscopy by designing the blinking behavior of targets with engineered binding frequency and duration in DNA-PAINT. We assay this kinetic barcoding approach in silico and in vitro using DNA origami structures, show the applicability for multiplexed RNA and protein detection in cells, and finally experimentally demonstrate 124-plex super-resolution imaging within minutes.
Keyphrases
- single molecule
- high resolution
- circulating tumor
- cell free
- label free
- nucleic acid
- induced apoptosis
- high throughput
- single cell
- loop mediated isothermal amplification
- high speed
- real time pcr
- mass spectrometry
- binding protein
- transcription factor
- endoplasmic reticulum stress
- optical coherence tomography
- signaling pathway
- photodynamic therapy
- cell proliferation
- cell death
- pi k akt
- molecular dynamics simulations