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Single-molecule Super-resolution Imaging of Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane with Novel Fluorescent Probes
Published on: October 15, 2016
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New Dual Fluorescent Probe for Simultaneous Biothiol and Phosphate Bioimaging
Sandra Resa1, Angel Orte2, Delia Miguel1
1Department of Organic Chemistry, Faculty of Sciences, University of Granada, C. U. Fuentenueva s/n, 18071 Granada (Spain), Fax: (+34) 958248437.
Chemistry (Weinheim an Der Bergstrasse, Germany)
|September 17, 2015
Summary
A novel dual fluorescent probe simultaneously detects biothiols and phosphate anions in cells. This advancement aids in understanding metabolite relationships in both healthy and diseased cells.
Area of Science:
- Chemical Biology
- Molecular Imaging
- Biochemistry
Background:
- Simultaneous metabolite detection offers insights into cellular health and disease.
- Fluorescent probes are crucial for in vivo biological analysis due to their low toxicity.
- Understanding the interplay of biothiols and phosphate anions is vital in cellular processes.
Purpose of the Study:
- To design and synthesize a novel dual fluorescent probe for simultaneous detection of biothiols and phosphate anions.
- To investigate the mechanism of action for the dual probe in biological systems.
- To validate the probe's efficacy in vitro and in live cells.
Main Methods:
- Synthesis of a 2,4-dinitrobenzenesulfinate (DNBS) derivative of Granada Green (GG).
- Characterization using steady-state and time-resolved fluorescence spectroscopy.
- Intracellular detection via fluorescence-lifetime imaging microscopy (FLIM) in HeLa cells.
Main Results:
- A new sulfinyl xanthene derivative (DNBS-GG) was successfully synthesized.
- The probe demonstrated simultaneous sensing of biological thiols and phosphate anions.
- The probe released fluorescent GG via thiolysis and responded to phosphate through fluorescence decay time.
- Successful intracellular imaging in HeLa cells was achieved.
Conclusions:
- The developed dual probe enables simultaneous detection of biothiols and phosphate anions.
- This probe is a valuable tool for studying metabolite dynamics in live cells.
- The findings open new avenues for understanding cellular signaling pathways.
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