Issue 8, 2023

Super-resolution imaging of linearized chromatin in tunable nanochannels

Abstract

Nanofluidic linearization and optical mapping of naked DNA have been reported in the research literature, and implemented in commercial instruments. However, the resolution with which DNA features can be resolved is still inherently limited by both Brownian motion and diffraction-limited optics. Direct analysis of native chromatin is further hampered by difficulty in electrophoretic manipulation, which is routinely used for DNA analysis. This paper describes the development of a three-layer, tunable, nanochannel system that enables non-electrophoretic linearization and immobilization of native chromatin. Furthermore, through careful selection of self-blinking fluorescent dyes and the design of the nanochannel system, we achieve direct stochastic optical reconstruction microscopy (dSTORM) super-resolution imaging of the linearized chromatin. As an initial demonstration, rDNA chromatin extracted from Tetrahymena is analyzed by multi-color imaging of total DNA, newly synthesized DNA, and newly synthesized histone H3. Our analysis reveals a relatively even distribution of newly synthesized H3 across two halves of the rDNA chromatin with palindromic symmetry, supporting dispersive nucleosome segregation. As a proof-of-concept study, our work achieves super-resolution imaging of native chromatin fibers linearized and immobilized in tunable nanochannels. It opens up a new avenue for collecting long-range and high-resolution epigenetic information as well as genetic information.

Graphical abstract: Super-resolution imaging of linearized chromatin in tunable nanochannels

Supplementary files

Article information

Article type
Communication
Submitted
15 Mar 2023
Accepted
16 May 2023
First published
24 May 2023

Nanoscale Horiz., 2023,8, 1043-1053

Author version available

Super-resolution imaging of linearized chromatin in tunable nanochannels

J. Lee, J. H. Chiu, N. J. Ginga, T. Ahmed, M. D. Thouless, Y. Liu and S. Takayama, Nanoscale Horiz., 2023, 8, 1043 DOI: 10.1039/D3NH00096F

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