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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.
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A specific probe for two-photon fluorescence lysosomal imaging.

Zhiguo Wu1, Mingliang Tang, Tian Tian

  • 1State Key Laboratory of Virology, College of Chemistry and Molecular Sciences, Wuhan University, Hubei, Wuhan, PR China.

Talanta
|November 22, 2011
PubMed
Summary
This summary is machine-generated.

Researchers developed a novel lysosome-specific probe, W-7, for advanced cellular imaging. This probe demonstrates high specificity and selectivity, enabling detailed visualization of lysosomes in living cells using two-photon microscopy.

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Area of Science:

  • Cell Biology
  • Chemical Biology
  • Biomedical Imaging

Background:

  • Lysosomes are crucial organelles involved in cellular degradation and are implicated in cancer progression.
  • Targeting and imaging lysosomes are essential for understanding their role in physiological and pathological processes.
  • Developing specific probes is key for accurate lysosome visualization and analysis.

Purpose of the Study:

  • To design and synthesize a novel lysosome-specific probe, W-7.
  • To evaluate the optical properties and two-photon absorption (TPA) characteristics of W-7.
  • To assess the specificity, selectivity, and imaging capabilities of W-7 in living cells.

Main Methods:

  • Synthesis of W-7 probe using one-pot reaction and Heck reaction.
  • Characterization of optical properties and TPA effect of W-7.
  • In vivo imaging experiments in HeLa, MRC-5, and NRK cells.
  • Comparison with existing probes (LT Red, GT Red, MT Red).
  • Two-photon fluorescence microscopy and 3D reconstruction.
  • Chiral-High Performance Liquid Chromatography (HPLC) for enantioseparation.

Main Results:

  • W-7 was successfully synthesized from Tröger's base.
  • W-7 exhibited two-photon absorption (TPA) under infrared laser excitation.
  • W-7 demonstrated high specificity and selectivity for lysosomes in various living cell lines (R=0.96).
  • High-resolution two-photon fluorescence images and 3D reconstructions of lysosomes were obtained.
  • The enantiomers of W-7 showed comparable performance in lysosome imaging.

Conclusions:

  • The novel probe W-7 is a highly specific and selective tool for lysosome targeting and imaging.
  • W-7 enables advanced visualization of lysosomes using two-photon microscopy in living cells.
  • The findings contribute to the development of new probes for studying lysosome function and related diseases.