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Grant support

This research was funded by the Science and Innovation Spanish Ministery (grant nos. RTI2018-101050-J-I00, PID2019-106211RB-I00, and EIN2020-112419. P.R.S. is grateful for a Juan de la Cierva Incorporacion scholarship (IJC2019-041915-I).

Analysis of institutional authors

Rodriguez-Sevilla, PCorresponding AuthorJaque, DAuthor

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Review

Multichannel Fluorescence Microscopy: Advantages of Going beyond a Single Emission

Publicated to:ADVANCED NANOBIOMED RESEARCH. 2 (5): 2100084- - 2022-05-01 2(5), DOI: 10.1002/anbr.202100084

Authors: Rodriguez-Sevilla, Paloma; Thompson, Sebastian A; Jaque, Daniel

Affiliations

Hosp Ramon & Cajal, Inst Ramon & Cajal Invest Sanitaria, Ctra Colmenar km 9 100, Madrid 28034, Spain - Author
Madrid Inst Adv Studies Nanosci, IMDEA Nanociencia, Faraday 9, Madrid 28049, Spain - Author
Natl Ctr Biotechnol CNB CSIC IMDEA, Nanobiotechnol Unit Associated, Madrid 28049, Spain - Author
Univ Autonoma Madrid, Dept Fis Mat, Nanomat Bioimaging Grp NanoBIG, Francisco Tomas & Valiente 7, Madrid 28049, Spain - Author

Abstract

Fluorescent microscopy has enabled the study of intracellular processes and revealed the most intricate details of the subcellular structure. This has benefitted not only the basic biological science, but also has had an impact in numerous biomedical applications. Basic fluorescent sensing techniques use the change in the absolute emission of a fluorescent sensor. This entails some disadvantages as the signal might be influenced by factors not directly related to the process under study (e.g., fluctuations in the excitation source). To overcome these drawbacks, one can use multiple emissions of a single or various fluorophores. There are numerous examples of multichannel fluorescence microscopy techniques that have given rise to numerous ratiometric methods and multiplexing assays. Herein, how the use of multiple emission channels has impacted fluorescence microscopy in terms of speed, sensitivity, and resolution is reviewed. Using recent examples, how the easy implementation of multichannel detection can overcome current limitations of the main used fluorescence techniques and promote the development of novel microscopy methods is shown.

Keywords

Biomedical applicationsDotsExpressionFluorescence microscopyFretImaging revealsIntracellular sensingLiving cellsMultichannelMultiplexingNanoparticlesProbes designQuantificationRatiometric detectionRatiometric sensingViscosity

Quality index

Bibliometric impact. Analysis of the contribution and dissemination channel

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: 1.81. 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 14, 2024)

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:

  • Field Citation Ratio (FCR) from Dimensions: 2.78 (source consulted: Dimensions Jun 2025)

Specifically, and according to different indexing agencies, this work has accumulated citations as of 2025-06-20, the following number of citations:

  • WoS: 9
  • Scopus: 12
  • OpenCitations: 11

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 2025-06-20:

  • 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: 26 (PlumX).

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.
  • Assignment of a Handle/URN as an identifier within the deposit in the Institutional Repository: https://repositorio.uam.es/handle/10486/714139

Leadership analysis of institutional authors

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 (RODRIGUEZ SEVILLA, PALOMA) and Last Author (JAQUE GARCIA, DANIEL).

the author responsible for correspondence tasks has been RODRIGUEZ SEVILLA, PALOMA.