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Updated: Jun 12, 2026
09:11
High-resolution Volume Imaging of Neurons by the Use of Fluorescence eXclusion Method and Dedicated Microfluidic Devices
Published on: March 26, 2018
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Confinement-sensitive volume regulation dynamics via high-speed nuclear morphological measurements.
Yixuan Li1, Hui Ting Ong1, Hongyue Cui1
1Mechanobiology Institute, National University of Singapore, 117411, Singapore.
Summary
Cellular confinement impacts nuclear volume regulation. New microscopy reveals cells rapidly decrease nuclear volume upon entering confinement, deviating from standard osmotic response models.
Area of Science:
- Cellular mechanics and biophysics
- Microfluidics and live-cell imaging
Background:
- Cells in vivo experience physical confinement, altering nuclear dynamics.
- Existing methods for studying nuclear deformation are limited by speed and phototoxicity.
- Understanding nuclear volume regulation in confined environments is crucial for cell migration studies.
Purpose of the Study:
- To develop a high-time-resolution imaging technique for tracking nuclear volume dynamics in confined migrating cells.
- To investigate how cells regulate nuclear volume when entering physiological confinement.
- To explore the nuclear envelope's role in volume regulation under confinement.
Main Methods:
- Developed double fluorescence exclusion microscopy for imaging at PDMS microchannel sidewalls.
- Estimated nuclear surface area by verifying vertical symmetry in confined nuclei.
- Tracked nuclear volume and surface area dynamics at >30 frames per minute.
- Utilized osmotic shock to probe nuclear volume response in confinement.
Main Results:
- Cells rapidly expand surface area upon entering confinement, followed by a decrease in nuclear volume.
- Nuclear volume regulation under confinement deviates from classical osmotic scaling laws.
- The nuclear envelope's limited expansion potential appears to constrain volume regulation in confined cells.
Conclusions:
- The developed microscopy technique enables high-resolution tracking of nuclear volume dynamics during confined migration.
- Nuclear volume regulation in confinement is a rapid, dynamic process distinct from unconfined responses.
- The nuclear envelope's mechanical properties play a critical role in limiting nuclear volume changes within confined spaces.
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