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pH01:24

pH

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The potential of hydrogen (pH) is a measure of the acidity or basicity of a water-based solution determined by the concentration of hydronium ions (H3O+). In one liter of pure water at neutral pH, there are 1×10−7 moles of hydronium ions. However, the extensive range of hydronium ion concentrations present in water-based solutions makes measuring pH in moles cumbersome. Therefore, a pH scale was developed to convert moles of hydronium ions into the negative logarithm of the hydronium...
139.9K

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Measuring Phagosome pH by Ratiometric Fluorescence Microscopy
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A Mitochondria-Targeted Ratiometric Fluorescent pH Probe.

Xiangjun Liu1,2, Linlin Wang1,2, Tao Bing1,2

  • 1Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, CAS Research/Education Center for Excellence in Molecular Sciences, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China.

ACS Applied Bio Materials
|January 13, 2022
PubMed
Summary

Researchers developed IR-ANNA, a novel fluorescent probe for tracking mitochondrial pH (pHm) in real-time. This tool offers accurate monitoring of pHm changes in living cells, crucial for understanding physiological processes.

Keywords:
cyaninemitochondrial targetingnaphthalimidepH imagingratiometric fluorescent probe

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Biomedical Engineering

Background:

  • Mitochondrial pH (pHm) is vital for cellular functions.
  • Accurate monitoring of pHm is essential for physiological studies.
  • Existing methods may have limitations in specificity or real-time monitoring.

Purpose of the Study:

  • To develop a novel mitochondria-targeted ratiometric fluorescent probe for monitoring pHm.
  • To evaluate the probe's targeting ability, pH responsiveness, and cytotoxicity.
  • To demonstrate the probe's utility in tracking pHm changes in living cells under various stimuli.

Main Methods:

  • Synthesis of a ratiometric fluorescent probe (IR-ANNA) by conjugating a pH-sensitive naphthalimide derivative (ANNA) with a mitochondria-targeted, pH-insensitive cyanine dye.
  • Assessment of mitochondria-targeting efficiency.
  • Evaluation of pH response and calibration.
  • Cytotoxicity assays in living cells.
  • Monitoring of pHm changes in response to chemical stimuli (FCCP, NAC, H2O2) in living cells.

Main Results:

  • The developed probe, IR-ANNA, exhibited excellent mitochondria-targeting capabilities.
  • IR-ANNA demonstrated a robust and sensitive response to pH variations.
  • The probe showed low cytotoxicity, ensuring its suitability for live-cell imaging.
  • Successful real-time monitoring of mitochondrial pH fluctuations induced by FCCP, NAC, and H2O2 was achieved in living cells.

Conclusions:

  • IR-ANNA is a highly effective fluorescent probe for real-time monitoring of mitochondrial pH.
  • Its specific targeting, sensitivity, and low toxicity make it a valuable tool for cellular physiology research.
  • This probe facilitates the study of mitochondrial dysfunction and related diseases.