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Related Experiment Video

Updated: Apr 19, 2026

High-resolution Volume Imaging of Neurons by the Use of Fluorescence eXclusion Method and Dedicated Microfluidic Devices

High-resolution Volume Imaging of Neurons by the Use of Fluorescence eXclusion Method and Dedicated Microfluidic Devices

Published on: March 26, 2018

Excluded-volume effects in living cells.

David Gnutt¹, Mimi Gao, Oliver Brylski

¹Department of Physical Chemistry II, Ruhr University Bochum, Universitätsstrasse 150, 44801 Bochum (Germany).

Angewandte Chemie (International Ed. in English)

|January 6, 2015

Summary

Cellular environments are crowded, but artificial crowding agents don't fully mimic the in-cell crowding effect. A new sensor reveals heterogeneous crowding that changes with cell stress, impacting biomolecular reactions.

Keywords:

FRET biophysics biosensors excluded-volume effect macromolecular crowding

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Area of Science:

Biophysics
Cell Biology
Biochemistry

Background:

Biomolecules function within crowded cellular environments.
Artificial macromolecular crowding agents are used to mimic cellular conditions in vitro.
The in-cell crowding effect remains unquantified, limiting understanding of its physicochemical properties.

Purpose of the Study:

To develop and utilize a sensor for quantifying macromolecular crowding effects within single living cells.
To compare in-cell crowding effects with those induced by artificial crowding agents.
To investigate the heterogeneity and dynamic changes of in-cell crowding.

Main Methods:

Development of a Förster Resonance Energy Transfer (FRET)-labeled polymer sensor.
Probing macromolecular crowding effects inside single living cells.
Analysis of sensor conformation under various cellular conditions, including cell stress.

Main Results:

Excluded-volume effects observed with artificial crowding agents do not compress the sensor in living cells.
The average sensor conformation in cells resembles that in aqueous buffer and cell lysate.
In-cell crowding is heterogeneous and significantly altered by cell stress.

Conclusions:

Artificial crowding agents do not fully replicate the in-cell crowding environment.
In-cell crowding is a dynamic and heterogeneous factor influencing biomolecular reactions.
The developed sensor provides a tool to systematically study in-cell crowding effects.