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Phase separation at the nanoscale quantified by dcFCCS.

Sijia Peng¹, Weiping Li¹, Yirong Yao¹

¹School of Life Sciences, Tsinghua-Peking Joint Center for Life Sciences, Beijing Advanced Innovation Center for Structural Biology, Beijing Frontier Research Center for Biological Structure, Tsinghua University, 100084 Beijing, China.

Proceedings of the National Academy of Sciences of the United States of America

|October 22, 2020

Summary

This summary is machine-generated.

Researchers discovered nanoscale biomolecular condensates using dual-color fluorescence cross-correlation spectroscopy (dcFCCS). This method quantifies their formation, size, and molecular interactions, revealing their importance in cellular processes.

Keywords:

condensate dual-color fluorescence cross-correlation spectroscopy fluorescence correlation spectroscopy liquid–liquid phase separation nanoscale

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Area of Science:

Biochemistry and Biophysics
Cell Biology
Molecular Interactions

Background:

Membraneless compartments, or biomolecular condensates, form via liquid-liquid phase separation and are crucial for cellular functions.
Conventional microscopy struggles to detect small, free-diffusing nanoscale condensates, leaving their formation and dynamics poorly understood.
Understanding the transition from soluble molecules to larger condensates is key to cellular regulation.

Purpose of the Study:

To investigate the formation and characteristics of nanoscale biomolecular condensates.
To overcome the limitations of conventional microscopy in detecting small condensates.
To quantify the properties of these nanoscale condensates and their constituent molecules.

Main Methods:

Development and application of dual-color fluorescence cross-correlation spectroscopy (dcFCCS).
Experimental measurements combined with Monte Carlo simulations.
Characterization of condensate size, growth rate, molecular stoichiometry, and binding affinity.

Main Results:

Successfully captured the formation of nanoscale condensates below the detection limit of conventional fluorescence microscopy.
Quantified key parameters of nanoscale condensates, including size, growth rate, and molecular composition.
Determined that the critical concentration for nanoscale condensate formation is significantly lower than previously detectable limits.

Conclusions:

Nanoscale biomolecular condensates form readily and play potentially significant roles in cellular processes.
Dual-color fluorescence cross-correlation spectroscopy (dcFCCS) is a powerful tool for studying these small condensates.
The study expands our understanding of phase separation dynamics and condensate biology at the nanoscale.