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Labeling DNA Probes03:31

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...

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A new rhodamine B-based lysosomal pH fluorescent indicator.

Hong-Shui Lv1, Shu-Ya Huang, Bao-Xiang Zhao

  • 1Institute of Organic Chemistry, School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100, PR China.

Analytica Chimica Acta
|July 13, 2013
PubMed
Summary

Researchers developed a new fluorescent probe, RCE, for accurately measuring acidic pH in cellular environments. This sensitive probe allows for real-time monitoring of pH changes in organelles like lysosomes within living cells.

Keywords:
Fluorescent probeIntracellular pHLiving cellsLysosomesRhodamine B

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Area of Science:

  • Chemical Biology
  • Cell Biology
  • Biochemistry

Background:

  • Accurate monitoring of intracellular pH is crucial for understanding cellular processes.
  • Existing pH probes may have limitations in sensitivity, selectivity, or photostability.

Purpose of the Study:

  • To design and synthesize a novel pH fluorescent probe, RCE, for precise detection of acidic environments.
  • To evaluate the probe's performance in terms of sensitivity, selectivity, response time, and applicability in living cells.

Main Methods:

  • Synthesis of the RCE probe based on rhodamine dye structural modifications.
  • Characterization of probe properties including fluorescence response, pKa, selectivity, and photostability.
  • In vitro and in vivo experiments to assess probe performance in cellular imaging and lysosomal pH monitoring.

Main Results:

  • The RCE probe demonstrated a >150-fold increase in fluorescence intensity within the pH range of 7.51-3.53.
  • The probe exhibited a pKa of 4.71, indicating suitability for studying acidic organelles.
  • RCE showed high selectivity, rapid response, low cytotoxicity, good photostability, reversibility, and cell membrane permeability.
  • Selective staining of lysosomes and monitoring of lysosomal pH changes in living cells were achieved.

Conclusions:

  • The RCE probe is a highly effective tool for real-time monitoring of acidic pH in biological systems.
  • Its favorable properties, including long wavelengths that minimize cell damage, make it valuable for studying acidic organelles like lysosomes.
  • RCE facilitates a deeper understanding of cellular pH dynamics and related biological processes.