Quantitative multiplexed imaging of chromatin ultrastructure with Decode-PAINT
biorxiv(2022)
Abstract
Super-resolution chromatin imaging techniques are emerging as powerful tools for spatial genomics. However, it remains challenging to visualize the genome on the order of tens of nanometers, a scale that can reveal rich structural detail about the nuclear organization. Here we introduce Decode-PAINT, a multiplexed in situ method, which leverages rapid diffraction-limited pre-decoding and DNA-PAINT (DNA points accumulation for imaging nanoscale topography). Decode-PAINT visualizes multiple discrete genomic regions at a resolution close to the nucleosome scale with a simple and easy-to-implement optical setup, reduces total imaging time by omitting multiple rounds of single-molecule recording, and enables the interrogation of the ultrastructure of the targets with <12 nm lateral and <20 nm axial resolution. We showcase the ability of our approach to perform multiplexed, high-resolution profiling of chromatin structure by performing targeted imaging of nine regions of active and inactive X chromosomes in individual cells. In short, Decode-PAINT accelerates in situ structural genomics.
### Competing Interest Statement
P.Y. is listed as an inventor for multiple patent and patent applications related to DNA-PAINT and SABER technology described in this paper. DNA-PAINT and SABER are licensed to Ultivue Inc. P.Y. is a co-founder, equity holder, director, and consultant of Ultivue Inc. J.Y.K. and B.J.B. hold patents on SABER.
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Key words
chromatin ultrastructure,quantitative multiplexed imaging,decode-paint
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