Imaging mass cytometry for high-dimensional tissue profiling in the eye.
IMC
Imaging mass cytometry
conjunctival melanoma
multi-dimensional cellular profiling
Journal
BMC ophthalmology
ISSN: 1471-2415
Titre abrégé: BMC Ophthalmol
Pays: England
ID NLM: 100967802
Informations de publication
Date de publication:
20 Sep 2021
20 Sep 2021
Historique:
received:
30
04
2021
accepted:
02
09
2021
entrez:
21
9
2021
pubmed:
22
9
2021
medline:
23
9
2021
Statut:
epublish
Résumé
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. 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. 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. 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.
Sections du résumé
BACKGROUND
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
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
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
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.
Identifiants
pubmed: 34544377
doi: 10.1186/s12886-021-02099-8
pii: 10.1186/s12886-021-02099-8
pmc: PMC8454101
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
338Informations de copyright
© 2021. The Author(s).
Références
Sci Rep. 2021 Jan 13;11(1):1023
pubmed: 33441834
Front Immunol. 2019 Nov 14;10:2657
pubmed: 31798587
Cytometry A. 2016 Mar;89(3):292-300
pubmed: 26355391
Br J Cancer. 1997;76(10):1353-60
pubmed: 9374383
Blood. 1993 Mar 15;81(6):1607-13
pubmed: 7680921
Angew Chem Int Ed Engl. 2007;46(32):6111-4
pubmed: 17533637
J Histochem Cytochem. 2000 May;48(5):653-62
pubmed: 10769049
Mol Imaging Biol. 2018 Dec;20(6):888-901
pubmed: 30167993
Can J Ophthalmol. 2019 Dec;54(6):699-707
pubmed: 31836103
Cell Syst. 2018 Jan 24;6(1):25-36.e5
pubmed: 29289569
Hum Pathol. 2020 Sep;103:107-119
pubmed: 32707054
Invest Ophthalmol Vis Sci. 1997 Jan;38(1):142-52
pubmed: 9008639
Ophthalmology. 1984 Jun;91(6):701-7
pubmed: 6205342
Oncotarget. 2017 May 20;8(33):54722-54734
pubmed: 28903377
Nat Methods. 2017 Sep;14(9):873-876
pubmed: 28783155
Genome Biol. 2006;7(10):R100
pubmed: 17076895
Cell Metab. 2019 Mar 5;29(3):755-768.e5
pubmed: 30713109
Nat Biotechnol. 2013 Jun;31(6):545-52
pubmed: 23685480
Anal Chem. 2009 Aug 15;81(16):6813-22
pubmed: 19601617
Nat Methods. 2014 Apr;11(4):417-22
pubmed: 24584193
J Cereb Blood Flow Metab. 2019 Mar;39(3):411-425
pubmed: 28933255
Front Neuroenergetics. 2010 May 21;2:
pubmed: 20725515
Cell. 2015 Jul 2;162(1):184-97
pubmed: 26095251
Cytometry A. 2017 Feb;91(2):160-169
pubmed: 28160444
Front Immunol. 2019 Oct 29;10:2534
pubmed: 31736961
Int J Mol Sci. 2020 Nov 30;21(23):
pubmed: 33266349
Semin Cancer Biol. 1990 Jun;1(3):199-206
pubmed: 2103495
Arch Ophthalmol. 2010 Feb;128(2):174-83
pubmed: 20142539
Mol Vis. 2011;17:1652-61
pubmed: 21738394
J Mol Diagn. 2012 Jan;14(1):22-9
pubmed: 22166544