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Quantification of Genetically Encoded Lipid Biosensors

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Phosphoinositides

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2251))

Abstract

Lipids, like phosphoinositides, can be visualized in living cells in real time using genetically encoded biosensors and fluorescence microscopy. Sensor localization can be quantified by determining the fluorescence intensity of each fluorophore. Enrichment of lipids at membranes can be determined by generating and applying an organelle-specific binary mask. In this chapter, we provide a detailed list of reagents and methods to visualize and quantify relative lipid levels. Applying this approach, changes in lipid levels can be assessed in cases when lipid metabolizing enzymes are mutated or otherwise altered.

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Acknowledgments

This work was supported by a National Institutes of Health grant (1R35GM119412-01) to G.R.V.H.

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Authors and Affiliations

  1. Department of Cell Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

    Rachel C. Wills, Jonathan Pacheco & Gerald R. V. Hammond

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  1. Rachel C. Wills
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  2. Jonathan Pacheco
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  3. Gerald R. V. Hammond
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Correspondence to Gerald R. V. Hammond .

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  1. Department of Chemistry and Biology and the Graduate Program in Molecular Science, Ryerson University, Toronto, ON, Canada

    Roberto J. Botelho

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Wills, R.C., Pacheco, J., Hammond, G.R.V. (2021). Quantification of Genetically Encoded Lipid Biosensors. In: Botelho, R.J. (eds) Phosphoinositides. Methods in Molecular Biology, vol 2251. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1142-5_4

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  • DOI: https://doi.org/10.1007/978-1-0716-1142-5_4

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