Evidence map›Paper›PMID 37801117›Full record

ArticleAnalytical and bioanalytical chemistry2024

Immunolabelling perturbs the endogenous and antibody-conjugated elemental concentrations during immuno-mass spectrometry imaging.

Monique G Mello, Mika T Westerhausen, Thomas E Lockwood, Prashina Singh, Jonathan Wanagat, David P Bishop

Open access · hybridAbstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Analytical and bioanalytical chemistry, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.

0numbers the graph read from it
0cells of the map it votes in
3citing papers in PubMed
0.9field-weighted citation impact, top 30% 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

3 citing papers in PubMed, 6 citations in OpenAlex.

  1. Review
  2. Article
  3. Article
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

6 authors at 2 institutions in 2 countries.

Monique G MelloHyphenated Mass Spectrometry Laboratory, Faculty of Science, University of Technology Sydney, P.O. Box 123, Broadway, NSW, 2007, Australia.
Mika T WesterhausenHyphenated Mass Spectrometry Laboratory, Faculty of Science, University of Technology Sydney, P.O. Box 123, Broadway, NSW, 2007, Australia.
Thomas E LockwoodHyphenated Mass Spectrometry Laboratory, Faculty of Science, University of Technology Sydney, P.O. Box 123, Broadway, NSW, 2007, Australia.
Prashina SinghHyphenated Mass Spectrometry Laboratory, Faculty of Science, University of Technology Sydney, P.O. Box 123, Broadway, NSW, 2007, Australia.
Jonathan Wanagat *Division of Geriatrics, Department of Medicine, David Geffen School of Medicine at UCLA, Los Angeles, CA, 90095, USA. JWanagat@mednet.ucla.edu.
David P Bishop *Hyphenated Mass Spectrometry Laboratory, Faculty of Science, University of Technology Sydney, P.O. Box 123, Broadway, NSW, 2007, Australia. david.bishop@uts.edu.au.
University of Technology Sydney · AUUniversity of California, Los Angeles · US

Funding

Mitochondrial DNA Deletion Mutation Frequency as a Metric of Biologic AgeR01AG069924 · NIA · UNIVERSITY OF CALIFORNIA LOS ANGELES · PI Jonathan Wanagat · 2022 to 2026
$2.1M
Mitochondrial biogenesis, genetics and cell loss in mammalian agingR01AG055518 · NIA · UNIVERSITY OF CALIFORNIA LOS ANGELES · PI AIKEN, JUDD M., HERBST, ALLEN · 2017 to 2022
$2.0M
Mitochondrial biogenesis, genetics and cell loss in mammalian agingK02AG059847 · NIA · UNIVERSITY OF CALIFORNIA LOS ANGELES · PI WANAGAT, JONATHAN · 2018 to 2022
$801k
Quantitative imaging of the dystrophin-glycoprotein complexR21AR072950 · NIAMS · UNIVERSITY OF CALIFORNIA LOS ANGELES · PI DOBLE, PHILIP, WANAGAT, JONATHAN · 2019 to 2020
$357k
NIAMS NIH HHS R21 AR072950NIA NIH HHS K02 AG059847NIA NIH HHS R01 AG055518NIA NIH HHS R01 AG069924NIH HHS K02AG059847NIH HHS R01AG055518NIH HHS R01AG069924
6 · The paper itself

Abstract

Immuno-mass spectrometry imaging uses lanthanide-conjugated antibodies to spatially quantify biomolecules via laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS). The multi-element capabilities allow for highly multiplexed analyses that may include both conjugated antibodies and endogenous metals to reveal relationships between disease and chemical composition. Sample handling is known to perturb the composition of the endogenous elements, but there has been little investigation into the effects of immunolabelling and coverslipping. Here, we used cryofixed muscle sections to examine the impact of immunolabelling steps on the concentrations of a Gd-conjugated anti-dystrophin primary antibody, and the endogenous metals Cu and Zn. Primary antibody incubation resulted in a decrease in Zn, and an increase in Cu. Zn was removed from the cytoplasm where it was hypothesised to be more labile, whereas concentrated locations of Zn remained in the cell membrane in all samples that underwent the immunostaining process. Cu increased in concentration and was found mostly in the cell membrane. The concentration of the Gd-conjugated antibody when compared to the standard air-dried sample was not significantly different when coverslipped using an organic mounting medium, whereas use of an aqueous mounting medium significantly reduced the concentration of Gd. These results build on the knowledge of how certain sample handling techniques change elemental concentrations and distributions in tissue sections. Immunolabelling steps impact the concentration of endogenous elements, and separate histological sections are required for the quantitative analysis of endogenous elements and biomolecules. Additionally, coverslipping tissue sections for complementary immunohistochemical/immunofluorescent imaging may compromise the integrity of the elemental label, and organic mounting media are recommended over aqueous mounting media.

Indexed as

Laser TherapyMetalsDiagnostic ImagingMass SpectrometryMetalsEndogenous elementsImmuno-mass spectrometry imagingLA-ICP-MS imaging

Identifiers

PMID37801117
PMCPMC10997740
OpenAlexW4387393191

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.