Evidence map›Paper›PMID 35163349›Full record

ReviewInternational journal of molecular sciences2022

Ten Approaches That Improve Immunostaining: A Review of the Latest Advances for the Optimization of Immunofluorescence.

Ricardo Piña, Alma I Santos-Díaz, Erika Orta-Salazar, Azucena Ruth Aguilar-Vazquez, Carola A Mantellero, Isabel Acosta-Galeana, Argel Estrada-Mondragon, Mara Prior-Gonzalez, Jadir Isai Martinez-Cruz, Abraham Rosas-Arellano

Open access · goldAbstract readReview
In one paragraph

Review in International journal of molecular sciences, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 61 papers.

0numbers the graph read from it
0cells of the map it votes in
61citing papers in PubMed
8.7field-weighted citation impact, top 1% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

61 citing papers in PubMed, 106 citations in OpenAlex.

  1. Review
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  8. Materials today. Bio · 2026
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  15. The Cytoskeleton in Space.Methods in molecular biology (Clifton, N.J.) · 2026
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  19. Estrogen-like Activity ofBiomedicines · 2025
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  20. Review

1 more citing papers are in PubMed but not listed here.

4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

10 authors at 5 institutions in 3 countries.

Ricardo PiñaDepartamento de Biología, Facultad de Ciencias Básicas, Universidad Metropolitana de Ciencias de la Educación, Santiago 7760197, Chile.ORCID 0000-0002-2316-8952
Alma I Santos-DíazDepartamento de Fisiología, Biofísica y Neurociencias, Cinvestav del IPN, Ciudad de México 07360, Mexico.
Erika Orta-SalazarDepartamento de Neurobiología del Desarrollo y Neurofisiología, Instituto de Neurobiología, Universidad Nacional Autónoma de México, Querétaro 76230, Mexico.
Azucena Ruth Aguilar-VazquezDepartamento de Neurobiología del Desarrollo y Neurofisiología, Instituto de Neurobiología, Universidad Nacional Autónoma de México, Querétaro 76230, Mexico.
Carola A MantelleroFacultad de Salud y Ciencias Sociales, Universidad de Las Américas, Santiago 7500975, Chile.
Isabel Acosta-GaleanaDivisión de Neurociencias, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Ciudad de México 04510, Mexico.
Argel Estrada-MondragonDepartment of Biomedical and Clinical Sciences, Linköping University, 581 83 Linköping, Sweden.
Mara Prior-GonzalezDivisión de Neurociencias, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Ciudad de México 04510, Mexico.
Jadir Isai Martinez-CruzDivisión de Neurociencias, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Ciudad de México 04510, Mexico.
Abraham Rosas-ArellanoUnidad de Imagenología, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Ciudad de México 04510, Mexico.
Universidad Nacional Autónoma de México · MXInstituto Politécnico Nacional · MXLinköping University · SEMetropolitan University of Educational Sciences · CLUniversidad Bernardo O'Higgins · CL

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Immunostaining has emerged as one of the most common and valuable techniques that allow the localization of proteins at a quantitative level within cells and tissues using antibodies coupled to enzymes, fluorochromes, or colloidal nanogold particles. The application of fluorochromes during immunolabeling is referred to as immunofluorescence, a method coupled to widefield or confocal microscopy and extensively applied in basic research and clinical diagnosis. Notwithstanding, there are still disadvantages associated with the application of this technique due to technical challenges in the process, such as sample fixation, permeabilization, antibody incubation times, and fluid exchange, etc. These disadvantages call for continuous updates and improvements to the protocols extensively described in the literature. This review contributes to protocol optimization, outlining 10 current methods for improving sample processing in different stages of immunofluorescence, including a section with further recommendations. Additionally, we have extended our own antibody signal enhancer method, which was reported to significantly increase antibody signals and is useful for cervical cancer detection, to improve the signals of fluorochrome-conjugated staining reagents in fibrous tissues. In summary, this review is a valuable tool for experienced researchers and beginners when planning or troubleshooting the immunofluorescence assay.

Indexed as

AntibodiesFluorescent DyesFluorescent Antibody TechniqueMicroscopy, ConfocalStaining and LabelingAntibodiesFluorescent Dyesantibodyimmunocytofluorescenceimmunohistofluorescenceimmunolabelingimmunolocalizationsignal-to-noise ratiounmask epitopes

Identifiers

PMID35163349
PMCPMC8836139
OpenAlexW4210511126

What OpenQuestion holds

Textmetadata
LicenceCC BY
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.