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Related Experiment Video

Updated: Jan 17, 2026

Single-molecule Super-resolution Imaging of Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane with Novel Fluorescent Probes
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Leaflet-specific phospholipid imaging using genetically encoded proximity sensors.

William M Moore1, Roberto J Brea1,2, Caroline H Knittel1

  • 1Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA, USA.

Nature Chemical Biology
|September 15, 2025
PubMed
Summary
This summary is machine-generated.

Researchers developed a new tool, FACES, to image cell lipids and their orientation in living cells. This method helps understand lipid transport and membrane asymmetry, advancing cell biology research.

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Area of Science:

  • Cell Biology
  • Biochemistry
  • Molecular Imaging

Background:

  • Cellular lipid composition is highly variable across organelles and membrane leaflets.
  • Existing tools lack the spatial resolution to measure lipid heterogeneity and transport in vivo.
  • Understanding lipid dynamics is crucial for cellular function and disease.

Purpose of the Study:

  • To develop a novel chemogenetic tool for quantitative imaging of subcellular lipid pools.
  • To determine the transbilayer orientation of lipids within living cells.
  • To investigate the roles of lipid transfer proteins and mechanisms generating membrane asymmetry.

Main Methods:

  • Developed fluorogen-activating coincidence encounter sensing (FACES), a tool combining bioorthogonal chemistry and genetically encoded fluorogen-activating proteins (FAPs).
  • Utilized FAPs for reversible proximity sensing of conjugated molecules.
  • Applied transmembrane domain-containing FAPs to assess lipid asymmetry in the trans-Golgi network.

Main Results:

  • FACES enables quantitative imaging of subcellular lipid pools and their transbilayer orientation in living cells.
  • Identified roles for lipid transfer proteins in trafficking phosphatidylcholine between the ER and mitochondria.
  • Demonstrated FACES's ability to reveal membrane asymmetry of multiple lipid classes and investigate its generation mechanisms.

Conclusions:

  • FACES is a versatile chemogenetic tool for in vivo lipid imaging and orientation analysis.
  • The study provides new insights into lipid transport and membrane asymmetry mechanisms.
  • FACES can be extended to image other molecule classes, such as glycans.