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Probing The Structure And Dynamics Of Nucleosomes Using Atomic Force Microscopy Imaging

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Published on: January 31, 2019

Single plane illumination microscopy as a tool for studying nucleome dynamics.

Jörg Langowski¹

¹Biophysics of Macromolecules, DKFZ Heidelberg, D-69120 Heidelberg, Germany.

Methods (San Diego, Calif.)

|June 27, 2017

Summary

Light sheet fluctuation microscopy analyzes dynamic molecular information using fluorescence intensity. This advanced optical method reveals insights into transcription factor dimerization, nuclear receptor activity, and chromatin viscoelasticity.

Keywords:

Fluorescence correlation spectroscopy Light sheet microscopy Nuclear architecture

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Area of Science:

Biophysics
Cell Biology
Microscopy

Background:

Single plane illumination microscopy (SPIM) offers high spatial resolution with reduced phototoxicity.
Fluorescence intensity fluctuations contain dynamic information about molecular behavior.
Understanding molecular dynamics is crucial for cell biology.

Purpose of the Study:

To review the principles of light sheet fluctuation microscopy.
To demonstrate applications in studying transcription factor and chromatin dynamics.
To elucidate molecular mechanisms underlying gene regulation and nuclear organization.

Main Methods:

Light sheet fluctuation microscopy (LSFM) utilizing fluorescence intensity fluctuations.
Application of LSFM to observe Fos and Jun protein dimerization.
Analysis of nuclear receptor dynamics and interphase chromatin viscoelasticity.

Main Results:

Fos and Jun protein dimerization is directly correlated with DNA binding.
Nuclear receptor activation significantly alters intranuclear dynamics.
Interphase chromatin viscoelasticity is highly dependent on lamin A presence.

Conclusions:

Light sheet fluctuation microscopy provides valuable insights into molecular dynamics.
LSFM elucidates the interplay between protein dimerization, DNA binding, and nuclear processes.
Lamin A plays a critical role in regulating chromatin mechanics.