Microchimica Acta

, 186:469 | Cite as

Ratiometric enhanced fluorometric determination and imaging of intracellular microRNA-155 by using carbon dots, gold nanoparticles and rhodamine B for signal amplification

  • Somayeh Hamd-Ghadareh
  • Baram Ahmed Hamah-Ameen
  • Abdollah SalimiEmail author
  • Fardin Fathi
  • Farzad Soleimani
Original Paper


An ultrasensitive and highly reliable ratiometric assay is described for the determination of microRNA-155. It works at the attomolar concentration level and has high selectivity which warrants its potential application in cancer biomarker tracking. The excellent performance of this method results from (a) the use of a hybrid conjugate prepared from Rhodamine B (RhB), carbon dots (CDs) and probe-microRNA, and (b) from the measurement of fluorescence resonance energy transfer (FRET) that is observed in the AuNP/target-microRNA system as a result of RNA hybridization. The dye RhB (emission peak at 580 nm) serves as an internal reference. The sensitivity of this assay is increased by about 30% because of the broad emissions of CDs (489 nm and 665 nm) through a sequential FRET phenomenon. RhB-CDs were covalently bio-conjugated to probe microRNA. In the presence of AuNPs, the fluorescence of the CDs is quenched, while in the presence of microRNA-155, the ratio of fluorescences at 489 and 665 nm (I489/I665) is enhanced again. A linear relationship exists between the ratio of fluorescence and the concentration of microRNA-155 in the range from 1 aM to 0.1 μM, and the detection limit is 0.3 aM. The assay was applied to quantitative studies of target microRNA-155 in multiple pathways associated with cancer progression in biological fluids include human serum samples and cancer cells. The nanoprobe also deliver clear signal to microRNA target in fixed and lived MDA-MB-231 cells.

Graphical abstract

A ratiometric FRET sensing method used for microRNA-155 detection at aM concentration level using CDs and AuNPs as donor–acceptor respectively and Rhodamine B as amplification reagent. The application of assay for imaging of microRNA-155 in fixed and live MDA-MB-231 cells is demonstrated.


Hybridization Live cell Ratiometric sensing Dual emission Fluorescence resonance energy transfer 



This research was supported by the Iranian Nanotechnology Initiative (142565) and the Research Offices of the University of Kurdistan (grant number 4.1404086).

Compliance with ethical standards

The author(s) declare that they have no competing interests.

Supplementary material

604_2019_3446_MOESM1_ESM.docx (2 mb)
ESM 1 (DOCX 1.95 mb)


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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  • Somayeh Hamd-Ghadareh
    • 1
    • 2
  • Baram Ahmed Hamah-Ameen
    • 2
  • Abdollah Salimi
    • 1
    • 3
    Email author
  • Fardin Fathi
    • 4
  • Farzad Soleimani
    • 4
  1. 1.Department of ChemistryUniversity of KurdistanSanandajIran
  2. 2.Charmo Research CenterCharmo UniversitySulaimaniIraq
  3. 3.Research Center for NanotechnologyUniversity of KurdistanSanandajIran
  4. 4.Cellular and Molecular Reserch CenterKurdistan University of Medical SciencesSanandajIran

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