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Highly Multiplexed, Super-resolution Imaging of T Cells Using madSTORM

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Mapping the nuclear landscape with multiplexed super-resolution fluorescence microscopy.

Fariha Rahman, Victoria Augoustides, Emma Tyler

Biorxiv : the Preprint Server for Biology

|August 30, 2024

Summary

This summary is machine-generated.

Researchers developed a novel imaging method to map nuclear organization. This technique reveals how active and repressive nuclear states are organized across different scales, offering new insights into cellular structure.

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Area of Science:

Cell Biology
Molecular Biology
Genomics

Background:

The cell nucleus orchestrates vital cellular functions.
Understanding spatial organization of nuclear components like DNA, protein complexes, and epigenetic marks is crucial.
Existing imaging techniques have limitations in resolving structures across multiple length scales.

Purpose of the Study:

To develop a multiplexed imaging protocol for high-precision localization of multiple nuclear targets.
To investigate the spatial organization of active and repressive nuclear states.
To explore the relationship between molecular organization and nuclear function.

Main Methods:

Developed a multiplexed imaging protocol.
Localized 13 different nuclear targets with nanometer precision in single cells.
Analyzed spatial correlations of nuclear components at micro- and nanoscale.

Main Results:

Nuclear states exist on a spectrum of length scales, with distinct nanoscale organizational principles.
Heterochromatin and euchromatin exhibit unique nanoscale organization.
Correlation of Heterochromatin protein 1-alpha (HP1-α) with DNA varied by scale.
RNA Polymerase II, p300, and CDK9 showed scale-dependent partitioning.
Perturbations in histone acetylation or transcription affected nanoscale organization.

Conclusions:

The developed imaging and analysis pipeline provides high-resolution insights into nuclear organization.
Nuclear organization principles differ significantly at the micro- and nanoscale.
This approach can be applied to study other cellular compartments.