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Measuring plasma membrane fluidity using confocal microscopy
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Measuring plasma membrane fluidity using confocal microscopy
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Journal Article
Measuring plasma membrane fluidity using confocal microscopy
2025
Overview
Membrane fluidity is a crucial parameter for cellular physiology. Recent evidence suggests that fluidity varies between cell types and states and in diseases. As membrane fluidity has gradually become an important consideration in cell biology and biomedicine, it is essential to have reliable and quantitative ways to measure it in cells. In the past decade, there has been substantial progress both in chemical probes and in imaging tools to make membrane fluidity measurements easier and more reliable. We have recently established a robust pipeline, using confocal imaging and new environment-sensitive probes, that has been successfully used for several studies. Here we present our detailed protocol for membrane fluidity measurement, from labeling to imaging and image analysis. The protocol takes ~4 h and requires basic expertise in cell culture, wet lab and microscopy.
Key points
This protocol describes a robust pipeline for measuring plasma membrane fluidity using confocal imaging and new environment-sensitive probes. Detailed steps cover the labeling and imaging of cells and the subsequent image analysis.
Compared with other methods for measuring membrane fluidity, this approach is easy to implement, as it can be performed with confocal microscopes and does not require a sophisticated data-analysis pipeline.
Membrane fluidity is a crucial feature in understanding cellular physiology. This protocol describes a robust pipeline for measuring plasma membrane fluidity using confocal imaging and new environment-sensitive probes.
Publisher
Nature Publishing Group UK,Nature Publishing Group
