Imaging mass cytometry for high-dimensional tissue profiling in the eye
Abstract Background Imaging mass cytometry (IMC) combines the principles of flow cytometry and mass spectrometry (MS) with laser scanning spatial resolution and offers unique advantages for the analysis of tissue samples in unprecedented detail. In contrast to conventional immunohistochemistry, whic...
Main Authors: | , , , , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
BMC
2021-09-01
|
Series: | BMC Ophthalmology |
Subjects: | |
Online Access: | https://doi.org/10.1186/s12886-021-02099-8 |
id |
doaj-73d3a0361d9c42be97daa3a33524a5ea |
---|---|
record_format |
Article |
spelling |
doaj-73d3a0361d9c42be97daa3a33524a5ea2021-09-26T11:56:15ZengBMCBMC Ophthalmology1471-24152021-09-012111910.1186/s12886-021-02099-8Imaging mass cytometry for high-dimensional tissue profiling in the eyeAnja Schlecht0Stefaniya Boneva1Henrike Salie2Saskia Killmer3Julian Wolf4Rozina Ida Hajdu5Claudia Auw-Haedrich6Hansjürgen Agostini7Thomas Reinhard8Günther Schlunck9Bertram Bengsch10Clemens AK Lange11Faculty of Medicine, Eye Center, University of FreiburgFaculty of Medicine, Eye Center, University of FreiburgFaculty of Medicine, Department of Medicine II, Gastroenterology, Hepatology, Endocrinology and Infectious Disease, University Medical Center FreiburgFaculty of Medicine, Department of Medicine II, Gastroenterology, Hepatology, Endocrinology and Infectious Disease, University Medical Center FreiburgFaculty of Medicine, Eye Center, University of FreiburgFaculty of Medicine, Eye Center, University of FreiburgFaculty of Medicine, Eye Center, University of FreiburgFaculty of Medicine, Eye Center, University of FreiburgFaculty of Medicine, Eye Center, University of FreiburgFaculty of Medicine, Eye Center, University of FreiburgFaculty of Medicine, Department of Medicine II, Gastroenterology, Hepatology, Endocrinology and Infectious Disease, University Medical Center FreiburgFaculty of Medicine, Eye Center, University of FreiburgAbstract Background Imaging mass cytometry (IMC) combines the principles of flow cytometry and mass spectrometry (MS) with laser scanning spatial resolution and offers unique advantages for the analysis of tissue samples in unprecedented detail. In contrast to conventional immunohistochemistry, which is limited in its application by the number of possible fluorochrome combinations, IMC uses isoptope-coupled antibodies that allow multiplex analysis of up to 40 markers in the same tissue section simultaneously. Methods In this report we use IMC to analyze formalin-fixed, paraffin-embedded conjunctival tissue. We performed a 18-biomarkers IMC analysis of conjunctival tissue to determine and summarize the possibilities, relevance and limitations of IMC for deciphering the biology and pathology of ocular diseases. Results Without modifying the manufacturer’s protocol, we observed positive and plausible staining for 12 of 18 biomarkers. Subsequent bioinformatical single-cell analysis and phenograph clustering identified 24 different cellular clusters with distinct expression profiles with respect to the markers used. Conclusions IMC enables highly multiplexed imaging of ocular samples at subcellular resolution. IMC is an innovative and feasible method, providing new insights into ocular disease pathogenesis that will be valuable for basic research, drug discovery and clinical diagnostics.https://doi.org/10.1186/s12886-021-02099-8Imaging mass cytometryIMCmulti-dimensional cellular profilingconjunctival melanoma |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Anja Schlecht Stefaniya Boneva Henrike Salie Saskia Killmer Julian Wolf Rozina Ida Hajdu Claudia Auw-Haedrich Hansjürgen Agostini Thomas Reinhard Günther Schlunck Bertram Bengsch Clemens AK Lange |
spellingShingle |
Anja Schlecht Stefaniya Boneva Henrike Salie Saskia Killmer Julian Wolf Rozina Ida Hajdu Claudia Auw-Haedrich Hansjürgen Agostini Thomas Reinhard Günther Schlunck Bertram Bengsch Clemens AK Lange Imaging mass cytometry for high-dimensional tissue profiling in the eye BMC Ophthalmology Imaging mass cytometry IMC multi-dimensional cellular profiling conjunctival melanoma |
author_facet |
Anja Schlecht Stefaniya Boneva Henrike Salie Saskia Killmer Julian Wolf Rozina Ida Hajdu Claudia Auw-Haedrich Hansjürgen Agostini Thomas Reinhard Günther Schlunck Bertram Bengsch Clemens AK Lange |
author_sort |
Anja Schlecht |
title |
Imaging mass cytometry for high-dimensional tissue profiling in the eye |
title_short |
Imaging mass cytometry for high-dimensional tissue profiling in the eye |
title_full |
Imaging mass cytometry for high-dimensional tissue profiling in the eye |
title_fullStr |
Imaging mass cytometry for high-dimensional tissue profiling in the eye |
title_full_unstemmed |
Imaging mass cytometry for high-dimensional tissue profiling in the eye |
title_sort |
imaging mass cytometry for high-dimensional tissue profiling in the eye |
publisher |
BMC |
series |
BMC Ophthalmology |
issn |
1471-2415 |
publishDate |
2021-09-01 |
description |
Abstract Background Imaging mass cytometry (IMC) combines the principles of flow cytometry and mass spectrometry (MS) with laser scanning spatial resolution and offers unique advantages for the analysis of tissue samples in unprecedented detail. In contrast to conventional immunohistochemistry, which is limited in its application by the number of possible fluorochrome combinations, IMC uses isoptope-coupled antibodies that allow multiplex analysis of up to 40 markers in the same tissue section simultaneously. Methods In this report we use IMC to analyze formalin-fixed, paraffin-embedded conjunctival tissue. We performed a 18-biomarkers IMC analysis of conjunctival tissue to determine and summarize the possibilities, relevance and limitations of IMC for deciphering the biology and pathology of ocular diseases. Results Without modifying the manufacturer’s protocol, we observed positive and plausible staining for 12 of 18 biomarkers. Subsequent bioinformatical single-cell analysis and phenograph clustering identified 24 different cellular clusters with distinct expression profiles with respect to the markers used. Conclusions IMC enables highly multiplexed imaging of ocular samples at subcellular resolution. IMC is an innovative and feasible method, providing new insights into ocular disease pathogenesis that will be valuable for basic research, drug discovery and clinical diagnostics. |
topic |
Imaging mass cytometry IMC multi-dimensional cellular profiling conjunctival melanoma |
url |
https://doi.org/10.1186/s12886-021-02099-8 |
work_keys_str_mv |
AT anjaschlecht imagingmasscytometryforhighdimensionaltissueprofilingintheeye AT stefaniyaboneva imagingmasscytometryforhighdimensionaltissueprofilingintheeye AT henrikesalie imagingmasscytometryforhighdimensionaltissueprofilingintheeye AT saskiakillmer imagingmasscytometryforhighdimensionaltissueprofilingintheeye AT julianwolf imagingmasscytometryforhighdimensionaltissueprofilingintheeye AT rozinaidahajdu imagingmasscytometryforhighdimensionaltissueprofilingintheeye AT claudiaauwhaedrich imagingmasscytometryforhighdimensionaltissueprofilingintheeye AT hansjurgenagostini imagingmasscytometryforhighdimensionaltissueprofilingintheeye AT thomasreinhard imagingmasscytometryforhighdimensionaltissueprofilingintheeye AT guntherschlunck imagingmasscytometryforhighdimensionaltissueprofilingintheeye AT bertrambengsch imagingmasscytometryforhighdimensionaltissueprofilingintheeye AT clemensaklange imagingmasscytometryforhighdimensionaltissueprofilingintheeye |
_version_ |
1716867632818290688 |