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Vincent Rigolot1, Christophe Biot1, Cedric Lion1

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Bioorthogonal chemistry enables probing biomolecules within cells using metabolic labeling. This review details challenges and strategies for imaging intracellular targets, overcoming limitations like reagent permeability and background fluorescence.

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Area of Science:

  • Chemical biology
  • Bioorthogonal chemistry
  • Molecular imaging

Background:

  • Bioorthogonal chemistry has revolutionized chemical biology over 20 years.
  • It enables probing biomolecules in biological fluids via metabolic labeling.
  • Key strategies include labeling glycans, proteins, and nucleic acids.

Purpose of the Study:

  • To review challenges in intracellular bioorthogonal chemistry.
  • To summarize strategies for imaging intracellular targets.
  • To address limitations in organelle-specific labeling.

Main Methods:

  • Discussing hurdles in intracellular bioorthogonal reactions.
  • Summarizing existing strategies for intracellular imaging.
  • Highlighting limitations such as membrane permeability and fluorescence background.

Main Results:

  • Bioorthogonal chemistry allows in situ biomolecule analysis.
  • Intracellular labeling faces challenges: reagent permeability, background fluorescence, and probe stability.
  • Mindful planning is crucial for successful intracellular imaging.

Conclusions:

  • Bioorthogonal chemistry offers powerful tools for chemical biology.
  • Imaging intracellular targets requires overcoming specific technical challenges.
  • This review provides a summary of strategies for successful intracellular bioorthogonal imaging.