Evidence map›Paper›PMID 39930005›Full record

ArticleScientific reports2025

Evaluation of methylene blue restaining versus conventional hydrogen peroxide decolorization in immunohistochemical diagnosis of melanoma.

Weisong Wan, Le Wang, Yuye Zeng, Yuchang Hu, Yufei Liu

Abstract readComparative Study
In one paragraph

Article in Scientific reports, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper, 1 of them a synthesis that pooled it.

0numbers the graph read from it
0cells of the map it votes in
1citing papers in PubMed, 1 pooled it
–field-weighted citation impact
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

1 citing paper in PubMed, 1 synthesis or guideline pooled it.

  1. Pooled it
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

5 authors.

Weisong WanThe First Clinical Medical College of China, Three Gorges University, Yichang, 443000, China.
Le WangThe First Clinical Medical College of China, Three Gorges University, Yichang, 443000, China.
Yuye ZengThe First Clinical Medical College of China, Three Gorges University, Yichang, 443000, China.
Yuchang HuThe First Clinical Medical College of China, Three Gorges University, Yichang, 443000, China. huyuchang@hotmail.com.
Yufei LiuThe First Clinical Medical College of China, Three Gorges University, Yichang, 443000, China. 402762573@qq.com.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

This study seeks to address the challenge of melanoma identification in immunohistochemical (IHC) diagnosis, which is complicated by the similar coloration of melanin and DAB (Diaminobenzidine) staining, by introducing methylene blue counterstaining as an innovative solution. We compared the effectiveness of methylene blue counterstaining with that of the traditional hydrogen peroxide bleaching method in the diagnosis of melanoma. The study included 46 paraffin-embedded melanoma samples, and the staining efficacy for markers such as Melan A, HMB-45, PRAME, and Ki-67 was assessed via both methods. The results demonstrated that methylene blue counterstaining effectively converted the brownish-yellow melanin granules to a deep green color, significantly enhancing contrast and clarity with DAB staining. The average contrast and clarity scores for the methylene blue counterstaining method were 1.96 ± 0.21 and 1.91 ± 0.28, respectively, which were significantly greater than those of the conventional IHC group and the hydrogen peroxide bleaching group (P < 0.01). Furthermore, methylene blue counterstaining did not cause noticeable tissue damage or cellular morphology distortion, with tissue integrity scores comparable to those of the conventional IHC group (P > 0.05). Although the contrast and clarity also improved in the hydrogen peroxide bleaching group, it resulted in a significant decrease in tissue integrity (P < 0.01). This study is the first to apply methylene blue counterstaining in melanoma IHC analysis, demonstrating its advantages in enhancing staining quality, simplifying procedural workflows, and preserving antigenicity. This method provides a novel and effective tool for the pathological diagnosis of melanoma, potentially improving diagnostic accuracy and reliability.

Indexed as

Hydrogen PeroxideImmunohistochemistryMelanomaMethylene BlueSkin NeoplasmsStaining and LabelingAdultAgedBiomarkers, TumorFemaleHumansMaleMiddle AgedBiomarkers, TumorHydrogen PeroxideMethylene BlueHydrogen peroxide bleachingImmunohistochemistryMelaninMelanomaMethylene blue restaining

Identifiers

PMID39930005
PMCPMC11811064

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Registered trials

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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.