Evidence map›Paper›PMID 33771067›Full record

ArticleThe Journal of international medical research2021

Optimizing tissue clearing and imaging methods for human brain tissue.

Min Sun Kim, Jang Ho Ahn, Ji Eun Mo, Ha Young Song, Deokhyeon Cheon, Seong Ho Yoo, Hyung Jin Choi

Abstract read
In one paragraph

Article in The Journal of international medical research, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.

0numbers the graph read from it
0cells of the map it votes in
4citing papers in PubMed
–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

4 citing papers in PubMed.

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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

7 authors.

Min Sun KimFunctional Neuroanatomy of Metabolism Regulation Laboratory, Department of Anatomy, Seoul National University College of Medicine, Seoul, South Korea.
Jang Ho AhnFunctional Neuroanatomy of Metabolism Regulation Laboratory, Department of Anatomy, Seoul National University College of Medicine, Seoul, South Korea.ORCID https://orcid.org/0000-0003-1058-812X
Ji Eun MoFunctional Neuroanatomy of Metabolism Regulation Laboratory, Department of Anatomy, Seoul National University College of Medicine, Seoul, South Korea.
Ha Young SongFunctional Neuroanatomy of Metabolism Regulation Laboratory, Department of Anatomy, Seoul National University College of Medicine, Seoul, South Korea.
Deokhyeon CheonFunctional Neuroanatomy of Metabolism Regulation Laboratory, Department of Anatomy, Seoul National University College of Medicine, Seoul, South Korea.
Seong Ho YooInstitute of Forensic Medicine and Department of Forensic Medicine, Seoul National University College of Medicine, Seoul, South Korea.
Hyung Jin ChoiFunctional Neuroanatomy of Metabolism Regulation Laboratory, Department of Anatomy, Seoul National University College of Medicine, Seoul, South Korea.ORCID https://orcid.org/0000-0003-0593-6978

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

objectivesTo identify optimum sample conditions for human brains, we compared the clearing efficiency, antibody staining efficiency, and artifacts between fresh and cadaver samples.

methodsFresh and cadaver samples were cleared using X-CLARITY™. Clearing efficiency and artifact levels were calculated using ImageJ, and antibody staining efficiency was evaluated after confocal microscopy imaging. Three staining methods were compared: 4-day staining (4DS), 11-day staining (11DS), and 4-day staining with a commercial kit (4DS-C). The optimum staining method was then selected by evaluating staining time, depth, method complexity, contamination, and cost.

resultsFresh samples outperformed cadaver samples in terms of the time and quality of clearing, artifacts, and 4',6-diamidino-2-phenylindole (DAPI) staining efficiency, but had a glial fibrillary acidic protein (GFAP) staining efficiency that was similar to that of cadaver samples. The penetration depth and DAPI staining improved in fresh samples as the incubation period lengthened. 4DS-C was the best method, with the deepest penetration. Human brain images containing blood vessels, cell nuclei, and astrocytes were visualized three-dimensionally. The chemical dye staining depth reached 800 µm and immunostaining depth exceeded 200 µm in 4 days.

conclusionsWe present optimized sample preparation and staining protocols for the visualization of three-dimensional macrostructure in the human brain.

Indexed as

BrainImaging, Three-DimensionalGlial Fibrillary Acidic ProteinHumansMicroscopy, ConfocalStaining and LabelingGlial Fibrillary Acidic Proteinconfocal microscopyDAPI stainingGFAP staininghuman brainTissue clearingX-CLARITY™

Identifiers

PMID33771067
PMCPMC8166401

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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.