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Temperature imaging using a cationic linear fluorescent polymeric thermometer and fluorescence lifetime imaging

Noriko Inada1, Nanaho Fukuda2, Teruyuki Hayashi3

  • 1Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Sakai, Japan. norikoinada@plant.osakafu-u.ac.jp.

Nature Protocols
|March 24, 2019
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Summary

Researchers developed a new method for imaging intracellular temperature in single cells using a fluorescent polymeric thermometer (FPT) and fluorescence lifetime imaging microscopy (FLIM). This technique offers high resolution for studying cellular thermal dynamics.

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Area of Science:

  • Cell Biology
  • Biophysics
  • Microscopy

Background:

  • Intracellular temperature is a critical physiological parameter for understanding cellular function.
  • Direct imaging of intracellular temperature at the single-cell level has been limited by the absence of suitable molecular thermometers.
  • Previous methods lacked the necessary resolution or applicability to live cells.

Purpose of the Study:

  • To develop and validate a novel method for high-resolution intracellular temperature imaging in live cells.
  • To assess the efficacy of a cationic linear fluorescent polymeric thermometer (FPT) for cellular thermometry.
  • To enable subcellular-level temperature distribution analysis.

Main Methods:

  • Utilized a cationic linear fluorescent polymeric thermometer (FPT) for its cell permeability and intracellular localization.
  • Employed fluorescence lifetime imaging microscopy (FLIM) for temperature measurements.
  • Developed a protocol integrating FPT incubation, calibration, and FLIM imaging, requiring approximately 14 hours.

Main Results:

  • The cationic linear FPT demonstrated rapid cell permeability (within 10 min) in various cell lines and yeast.
  • Achieved high temperature resolution (0.3–1.29 °C) within the physiological range (25–35 °C).
  • Enabled high spatial resolution (200 nm) for subcellular temperature mapping due to diffuse FPT localization.

Conclusions:

  • The developed FPT and FLIM-based method provides a powerful tool for live-cell intracellular thermometry.
  • This technique allows for unprecedented visualization of temperature dynamics at the subcellular level.
  • The protocol is suitable for researchers with experience in cell culture and confocal microscopy.