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Three-dimensional Optical-resolution Photoacoustic Microscopy
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Calcium Sensor for Photoacoustic Imaging.

Sheryl Roberts, Markus Seeger, Yuanyuan Jiang

  • 1CSSB, 22607 Hamburg and Institute for Biophysical Chemistry, Hannover Medical School , 30625 Hannover, Germany.

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|September 26, 2017
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We developed a new calcium sensor for photoacoustics (CaSPA) enabling live cell imaging. This tool allows visualization of calcium ion (Ca2+) fluxes in various biological systems with deep tissue penetration.

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Area of Science:

  • Biomedical Engineering
  • Optical Imaging
  • Molecular Biology

Background:

  • Photoacoustic imaging offers deep tissue penetration for mapping photoabsorbing molecules.
  • Existing calcium sensors lack selectivity or are not suitable for photoacoustic applications.
  • Understanding dynamic calcium ion (Ca2+) changes is crucial in cellular processes.

Purpose of the Study:

  • To develop a novel, cell-permeable calcium sensor optimized for photoacoustic imaging (CaSPA).
  • To enable real-time visualization of intracellular calcium ion (Ca2+) dynamics.
  • To demonstrate the sensor's utility in various biological models.

Main Methods:

  • Synthesis of a selective metallochromic sensor with specific optical properties (high extinction coefficient, low quantum yield, photobleaching resistance).
  • Optimization of the sensor for Ca2+-dependent photoacoustic signal changes, including a blueshift in absorbance.
  • Development of an acetoxymethyl esterified variant for efficient cell uptake and minimal toxicity.
  • Combined fluorescence and photoacoustic imaging in genetically unmodified cells, heart organoids, and zebrafish larval brain.

Main Results:

  • CaSPA exhibits a robust blueshift in absorbance upon binding micromolar concentrations of Ca2+.
  • Ca2+ binding leads to a decrease in the peak photoacoustic signal.
  • The esterified CaSPA variant demonstrated effective cell permeability and lack of toxicity.
  • Successful live imaging of Ca2+ fluxes was achieved in various biological systems, including complex tissues.

Conclusions:

  • CaSPA is a versatile and effective sensor for photoacoustic imaging of calcium ion (Ca2+) dynamics.
  • This technology facilitates unprecedented live imaging of Ca2+ fluxes in genetically unmodified biological models.
  • CaSPA opens new avenues for studying calcium signaling in health and disease.