Affordable Access

Access to the full text

In Vitro and in Cellulo Sensing of Transition Metals Using Time-Resolved Fluorescence Spectroscopy and Microscopy

Authors
  • Pal, Robert1
  • Barker, Abigail C. J.1
  • Hummel, Daniel1
  • Pålsson, Lars-Olof1
  • 1 Durham University, Department of Chemistry, Lower Mountjoy, Stockton Road, Durham, DH1 3LE, UK , Durham (United Kingdom)
Type
Published Article
Journal
Journal of Fluorescence
Publisher
Springer-Verlag
Publication Date
Dec 26, 2018
Volume
29
Issue
1
Pages
255–263
Identifiers
DOI: 10.1007/s10895-018-2335-z
Source
Springer Nature
Keywords
License
Green

Abstract

In this work we demonstrate that time domain techniques can be used successfully to monitor realtively weak modulations of the fluorescence in sensing applications. The metal sensing complex Newport Green DCF™ can detect selected transition metals in vivo as well as in vitro. Incremental addition of Ni and/or Zn (in vitro) lead to a substantial reduction in the yield of the fast component in a bi-exponential fluorescence decay (τ1 = 150–250 ps) from 60% to 30–35%. This is rationalised as an inhibition of intra-molecular electron transfer in the NPG sensing complex due to metal complexation. In order to explore this effect in cellulo, NIH 3 T3 mouse skin fibroplast cells were pre-incubated with set levels of Ni and Zn, at a constant concentration of NPG. The fluorescence modulation in cellullo was subsequently studied employing both time-resolved fluorescence microscopy and confocal fluorescence microscopy. In correlation with the in vitro observations, similar effects were observed on the fluorescence decay in cellulo.

Report this publication

Statistics

Seen <100 times