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Updated: Sep 15, 2025

Determination of Plasma Membrane Partitioning for Peripherally-associated Proteins
11:11

Determination of Plasma Membrane Partitioning for Peripherally-associated Proteins

Published on: June 15, 2018

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A gentle palette of plasma membrane dyes

Jing Ling1,2, Yitong Liu1,2, Alexandre Dumoulin3,4

  • 1Peking-Tsinghua Center for Life Science, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing 100871, China.

Proceedings of the National Academy of Sciences of the United States of America
|July 15, 2025
PubMed

Insights

New PK Mem dyes significantly reduce phototoxicity and enhance photostability for plasma membrane imaging. These advanced fluorescent probes enable ultralong-term live-cell imaging and superresolution microscopy of cellular dynamics.

Area of Science:

  • Cell Biology
  • Microscopy
  • Biochemistry

Background:

  • Plasma membrane (PM) stains are crucial for observing membrane structure and behavior.
  • Existing PM stains can cause photodynamic damage during prolonged fluorescence microscopy, limiting dynamic studies.
  • Need for improved dyes that are photostable and less toxic for live-cell imaging.

Purpose of the Study:

  • Introduce novel PK Mem dyes for advanced time-lapse fluorescence imaging of the plasma membrane.
  • Enhance photostability and reduce phototoxicity compared to existing membrane dyes.
  • Enable high-resolution imaging of cellular dynamics in various cell types and conditions.

Main Methods:

  • Development of PK Mem dyes by incorporating triplet-state quenchers into MemBright cyanine dyes with zwitterion anchors.
  • Testing dye performance in 2D and 3D imaging of live and fixed cancer cell lines and primary cells.
  • Evaluation of dye compatibility with superresolution techniques (STED/SIM) and in vivo imaging.

Main Results:

  • PK Mem dyes demonstrated a three-fold reduction in phototoxicity and over a four-fold increase in photostability.
  • Successful imaging of diverse cell types, including neurons, cardiomyocytes, and sperm cells.
  • Enabled ultralong-term recordings of cellular processes like cell migration and axonal growth cone dynamics.
  • Demonstrated compatibility with STED/SIM imaging for nanoscopic resolution of cellular events.

Conclusions:

  • PK Mem dyes offer a superior, gentle alternative for plasma membrane staining in time-lapse and superresolution microscopy.
  • These dyes facilitate the study of dynamic cellular processes previously limited by phototoxicity.
  • The enhanced photostability and reduced toxicity open new avenues for long-term live-cell imaging and nanoscale investigations.
Keywords:
long-term imagingmembrane dyesneuronal imagingphototoxicitysuper-resolution imaging

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