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Single-cell temperature mapping with fluorescent thermometer nanosheets.

Kotaro Oyama1,2,3,4, Mizuho Gotoh3,4, Yuji Hosaka1

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Cellular temperature measurements are unreliable. New fluorescent nanosheets offer noninvasive temperature mapping, revealing localized heat gradients rather than global changes in single cells.

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Area of Science:

  • Cellular Biology
  • Biophysics
  • Thermometry

Background:

  • Intracellular thermometers show heterogeneous cell temperatures (~1°C).
  • Previous methods face challenges excluding nonthermal factors, questioning reliability.
  • Accurate external measurement of cellular temperature is needed.

Purpose of the Study:

  • Develop a noninvasive method for global temperature mapping of single cells.
  • Investigate cellular temperature changes using novel fluorescent thermometer nanosheets.
  • Validate the accuracy and reliability of external cellular thermometry.

Main Methods:

  • Cultured cells (HeLa/HEK293, adipocytes, cardiomyocytes, neurons) on fluorescent nanosheets.
  • Utilized europium (III) thenoyltrifluoroacetonate trihydrate dye for temperature sensing.
  • Applied various stimuli including ionomycin, mitochondrial uncouplers, and electrical stimulation.

Main Results:

  • External measurements showed minimal temperature changes (<0.2°C) in nonexcitable cells.
  • Mutations causing Ca2+ leak did not significantly alter cellular temperature (<0.1°C).
  • Intracellular thermometry detected localized ER heat (~2°C), while global temperature remained stable (±0.2°C).
  • Excitable cells (cardiomyocytes, neurons) exhibited stable global temperatures during stimulation (±0.01-0.03°C).

Conclusions:

  • Heat production upon cell activation creates local temperature gradients (~1°C), not uniform global changes.
  • Novel nanosheet thermometry provides accurate, noninvasive cellular temperature monitoring.
  • Findings challenge previous intracellular thermometry interpretations and highlight localized thermal dynamics.