{rfName}
In

Indexed in

License and Use

Icono OpenAccess

Altmetrics

Impact on the Sustainable Development Goals (SDGs)

Analysis of institutional authors

Frances-Soriano, LauraAuthor

Share

August 7, 2025
Publications
>
Article

In Situ Rolling Circle Amplification Forster Resonance Energy Transfer (RCA-FRET) for Washing-Free Real-Time Single-Protein Imaging

Publicated to: Analytical Chemistry. 93 (3): 1842-1850 - 2021-01-26 93(3), DOI: 10.1021/acs.analchem.0c04828

Authors:

Frances-Soriano, Laura; Leino, Mattias; Dos Santos, Marcelina Cardoso; Kovacs, Daniel; Borbas, K Eszter; Soderberg, Ola; Hildebrandt, Niko
[+]

Affiliations

Univ Paris Saclay, Inst Integrat Biol Cell I2BC, CEA, CNRS, F-91405 Orsay, France - Author
Univ Rouen Normandie, Lab COBRA Chim Organ Bioorgan Reactivite & Anal, INSA, CNRS,NanoFRET Com, F-76821 Mont St Aignan, France - Author
Uppsala Univ, Dept Chem, Angstrom Lab, S-75120 Uppsala, Sweden - Author
Uppsala Univ, Dept Pharmaceut Biosci, S-75124 Uppsala, Sweden - Author
See more

Abstract

Fluorescence signal enhancement via isothermal nucleic acid amplification is an important approach for sensitive imaging of intra- or extracellular nucleic acid or protein biomarkers. Rolling circle amplification (RCA) is frequently applied for fluorescence in situ imaging but faces limitations concerning multiplexing, dynamic range, and the required multiple washing steps before imaging. Here, we show that Forster resonance energy transfer (FRET) between fluorescent dyes and between lanthanide (Ln) complexes and dyes that hybridize to beta-actin-specific RCA products in HaCaT cells can afford washing-free imaging of single beta-actin proteins. Proximity-dependent FRET could be monitored directly after or during (real-time monitoring) dye or Ln DNA probe incubation and could efficiently distinguish between photoluminescence from beta-actin-specific RCA and DNA probes freely diffusing in solution or nonspecifically attached to cells. Moreover, time-gated FRET imaging with the Ln-dye FRET pairs efficiently suppressed sample autofluorescence and improved the signal-to-background ratio. Our results present an important proof of concept of RCA-FRET imaging with a strong potential to advance in situ RCA toward easier sample preparation, higher-order multiplexing, autofluorescence-free detection, and increased dynamic range by real-time monitoring of in situ RCA.
[+]

Keywords

Affordable and clean energyAutofluorescenceCell lineComplexesDiagnostic assayDnDna probesFluorescence resonance energy transferFluorescent dyesHumansMicrornaNucleic acid amplification techniquesPlatformProteinsReductionTime factors

Quality index

Bibliometric impact. Analysis of the contribution and dissemination channel

The work has been published in the journal ANALYTICAL CHEMISTRY due to its progression and the good impact it has achieved in recent years, according to the agency WoS (JCR), it has become a reference in its field. In the year of publication of the work, 2021, it was in position 7/87, thus managing to position itself as a Q1 (Primer Cuartil), in the category Chemistry, Analytical. Notably, the journal is positioned above the 90th percentile.

From a relative perspective, and based on the normalized impact indicator calculated from World Citations provided by WoS (ESI, Clarivate), it yields a value for the citation normalization relative to the expected citation rate of: 1.66. This indicates that, compared to works in the same discipline and in the same year of publication, it ranks as a work cited above average. (source consulted: ESI Nov 13, 2025)

This information is reinforced by other indicators of the same type, which, although dynamic over time and dependent on the set of average global citations at the time of their calculation, consistently position the work at some point among the top 50% most cited in its field:

  • Weighted Average of Normalized Impact by the Scopus agency: 1.44 (source consulted: FECYT Mar 2025)

Specifically, and according to different indexing agencies, this work has accumulated citations as of 2026-04-06, the following number of citations:

  • WoS: 25
  • Scopus: 26
  • Europe PMC: 15
[+]

Impact and social visibility

From the perspective of influence or social adoption, and based on metrics associated with mentions and interactions provided by agencies specializing in calculating the so-called "Alternative or Social Metrics," we can highlight as of 2026-04-06:

  • The use, from an academic perspective evidenced by the Altmetric agency indicator referring to aggregations made by the personal bibliographic manager Mendeley, gives us a total of: 20.
  • The use of this contribution in bookmarks, code forks, additions to favorite lists for recurrent reading, as well as general views, indicates that someone is using the publication as a basis for their current work. This may be a notable indicator of future more formal and academic citations. This claim is supported by the result of the "Capture" indicator, which yields a total of: 20 (PlumX).

With a more dissemination-oriented intent and targeting more general audiences, we can observe other more global scores such as:

  • The Total Score from Altmetric: 3.

It is essential to present evidence supporting full alignment with institutional principles and guidelines on Open Science and the Conservation and Dissemination of Intellectual Heritage. A clear example of this is:

  • The work has been submitted to a journal whose editorial policy allows open Open Access publication.
Continuing with the social impact of the work, it is important to emphasize that, due to its content, it can be assigned to the area of interest of ODS 7 - Ensure access to affordable, reliable, sustainable and modern energy for all, with a probability of 87% according to the mBERT algorithm developed by Aurora University.
[+]

Leadership analysis of institutional authors

This work has been carried out with international collaboration, specifically with researchers from: France; Sweden.

There is a significant leadership presence as some of the institution’s authors appear as the first or last signer, detailed as follows: First Author (Francés Soriano, Laura) .

[+]

Project objectives

La aportación persigue los siguientes objetivos: analizar la aplicación de la amplificación en círculo rodante (RCA) combinada con transferencia de energía por resonancia de Förster (FRET) para la imagen en tiempo real de proteínas individuales sin necesidad de lavados; evaluar la capacidad de FRET para distinguir señales específicas de beta-actina en células HaCaT frente a señales no específicas; caracterizar la eficiencia de la imagen FRET con pares Ln-dye para suprimir la autofluorescencia y mejorar la relación señal-ruido; determinar el potencial de esta técnica para facilitar la preparación de muestras, aumentar la multiplexación y ampliar el rango dinámico mediante el monitoreo en tiempo real; y demostrar un concepto innovador que avance la imagen in situ basada en RCA hacia aplicaciones más sensibles y específicas.
[+]

Most relevant results

El estudio presenta avances significativos en la amplificación isotérmica in situ para la imagen de proteínas individuales mediante RCA-FRET. Se logró la detección sin lavado de proteínas beta-actina específicas en células HaCaT mediante transferencia de energía por resonancia de Förster (FRET) entre tintes fluorescentes y complejos de lantánidos. Se demostró la monitorización en tiempo real de FRET durante la incubación de sondas, permitiendo distinguir eficazmente la fotoluminiscencia específica de RCA de señales no específicas. La imagen con FRET temporizado suprimió eficazmente la autofluorescencia, mejorando la relación señal-ruido. Estos resultados evidencian un avance hacia una preparación de muestras más sencilla, mayor multiplexación, detección libre de autofluorescencia y un rango dinámico ampliado.
[+]

Awards linked to the item

We thank Lumiphore, Inc. for gifting Lumi4 reagents. This work was partially funded by the European Commission (Marie Sklodowska-Curie Actions, Individual Fellowship of Laura Frances-Soriano), the French National Research Agency (ANR projects "PhenX" and "Neutrinos" and Labex "SynOrg"), the Region Normandie (Chaire d'Excellence "COBRA-FRET"), the Institut Universitaire de France (IUF), the Swedish Research Council, and the Swedish Cancer Foundation.
[+]