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Fluorogenic Probes for Multicolor Imaging in Living Cells.

Gražvydas Lukinavičius1,2, Luc Reymond1, Keitaro Umezawa3

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Summary
This summary is machine-generated.

Researchers developed a new far-red fluorophore, SiR700, for advanced live-cell imaging. This silicon-rhodamine dye enables multicolor superresolution microscopy, enhancing cellular visualization capabilities.

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Area of Science:

  • Cellular and Molecular Imaging
  • Biophotonics
  • Organic Chemistry

Background:

  • Live-cell imaging requires fluorescent probes with specific spectral properties and compatibility with advanced microscopy techniques.
  • Existing probes may have limitations in spectral range or multicolor capabilities for superresolution applications.
  • Silicon-rhodamine dyes offer tunable optical properties suitable for biological imaging.

Purpose of the Study:

  • To introduce a novel far-red silicon-rhodamine fluorophore (SiR700) for live-cell multicolor imaging.
  • To develop and validate SiR700-based probes for various cellular targets and applications.
  • To demonstrate the utility of SiR700 in multicolor live-cell superresolution microscopy.

Main Methods:

  • Synthesis and characterization of the silicon-rhodamine-based fluorophore SiR700.
  • Development of SiR700-conjugated probes for F-actin, microtubules, lysosomes, and SNAP-tag.
  • Testing probe performance in live cells, including cell permeability, fluorogenicity, and compatibility with superresolution microscopy.
  • Performing multicolor imaging experiments in conjunction with SiR650-based probes.

Main Results:

  • SiR700 exhibits excitation and emission maxima at 690 nm and 715 nm, respectively.
  • SiR700-based probes are fluorogenic, cell-permeable, and suitable for superresolution microscopy.
  • Multicolor live-cell superresolution imaging was achieved in the far-red spectrum using SiR700 and SiR650 probes.
  • The new probes enable enhanced visualization of cellular structures and processes.

Conclusions:

  • SiR700 is a versatile far-red fluorophore for advanced live-cell imaging.
  • SiR700-based probes significantly expand the capacity for multicolor live-cell superresolution microscopy.
  • This development offers new possibilities for studying dynamic cellular processes with higher resolution and spectral multiplexing.