Abstract
It is now widely appreciated that the spatial organization of the genome is nonrandom, and its complex 3D folding has important consequences for many genome processes. Recent developments in multiplexed, super-resolution microscopy have enabled an unprecedented view of the polymeric structure of chromatin - from the loose folds of whole chromosomes to the detailed loops of cis-regulatory elements that regulate gene expression. Facilitated by the use of robotics, microfluidics, and improved approaches to super-resolution, thousands to hundreds of thousands of individual cells can now be analyzed in an individual experiment. This has led to new insights into the nature of genomic structural features identified by sequencing, such as topologically associated domains (TADs), and the nature of enhancer-promoter interactions underlying transcriptional regulation. We review these recent improvements.
🔬 Techniques
🧬 Organisms
✨ Fluorophores
🧪 Sample Preparation
🏛️ Research Organizations (ROR)
Affiliated research institutions:
📊 Figures
Figure 1.
Improvements in labeling and imaging chromatin.
A. Schematic of evolution in probes targeting the BX-C from Drosophila melanogaster over the last decade, taken from our own experiments. Changes in probe cost and production time are noted. B. Exampl...
Figure 2.
Similarities and differences explained: Diffraction-limited, STORM imaging, OligoSTORM, and ORCA.
A. in STORM (e.g. [ 14 ]) a sparse subset of fluors are activated at a time, so that their detected diffraction patterns (a.k.a. point-spread functions) donu2019t overlap. The centroids can be fit com...
Figure 3.
Limits of STORM.
For STORM (stochastic localization of individual fluors) to be advantageous, the absolute spread of the fluorescent molecules positions (purple stars) should be greater than the precision at which ind...
Figure 4.
Key Figure. Recent biological insights from advances in super-resolution imaging of chromatin.
A. Epigenetic domains are generally not resolved using conventional microscopy but reveal structurally distinct organization such as compaction, asphericity, and lack-of-mixing when resolved in super-...
Figure images are served from the NIH/NLM PubMed Central Open Access Subset or Europe PMC; copyright remains with the publishers and authors.
💬 Discussion
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