Measuring plasma membrane fluidity using spectral imaging

Membrane fluidity is a crucial parameter for cellular physiology. Recent evidence suggests that it varies in cell types, states, and in diseases. As membrane fluidity is gradually becoming a new player in cell biology and biomedicine, it is essential to have reliable and quantitative ways to measure it in cells.

In the last decade, there has been significant 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 spectral imaging and new environment-sensitive probes, which has been successfully used for several studies. In this protocol, we will present the key points of the membrane fluidity measurement from labelling to imaging and image analysis.

The protocol takes around 4 hours and it requires basic expertise in cell culture, wet lab and microscopy. Data usage Researchers are welcome to use the data contained in the dataset for any projects. Please cite this item upon use or when published.

We encourage reuse using the same CC BY 4.0 License. Data Content Excel files for graphs lsm files for raw microscopy images tif files for processed microscopy images svg files for figures Software to open files .csv - Microsoft Excel .lsm, .tif - Fiji (imagej.net/Fiji.html#Downloads) .svg - Inkscape