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Comparing and Correcting Spectral Sensitivities between Multispectral Microscopes: A Prerequisite to Clinical Implementation.
Eminizer, Margaret; Nagy, Melinda; Engle, Elizabeth L; Soto-Diaz, Sigfredo; Jorquera, Andrew; Roskes, Jeffrey S; Green, Benjamin F; Wilton, Richard; Taube, Janis M; Szalay, Alexander S.
Afiliação
  • Eminizer M; Department of Physics and Astronomy, Johns Hopkins University, Baltimore, MD 21210, USA.
  • Nagy M; Institute for Data Intensive Engineering and Science, Johns Hopkins University, Baltimore, MD 21210, USA.
  • Engle EL; Department of Physics and Astronomy, Johns Hopkins University, Baltimore, MD 21210, USA.
  • Soto-Diaz S; Institute for Data Intensive Engineering and Science, Johns Hopkins University, Baltimore, MD 21210, USA.
  • Jorquera A; Department of Dermatology, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
  • Roskes JS; Bloomberg-Kimmel Institute for Cancer Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
  • Green BF; Mark Foundation Center for Advanced Genomics and Imaging, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
  • Wilton R; Department of Dermatology, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
  • Taube JM; Bloomberg-Kimmel Institute for Cancer Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
  • Szalay AS; Mark Foundation Center for Advanced Genomics and Imaging, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
Cancers (Basel) ; 15(12)2023 Jun 08.
Article em En | MEDLINE | ID: mdl-37370719
ABSTRACT
Multispectral, multiplex immunofluorescence (mIF) microscopy has been used to great effect in research to identify cellular co-expression profiles and spatial relationships within tissue, providing a myriad of diagnostic advantages. As these technologies mature, it is essential that image data from mIF microscopes is reproducible and standardizable across devices. We sought to characterize and correct differences in illumination intensity and spectral sensitivity between three multispectral microscopes. We scanned eight melanoma tissue samples twice on each microscope and calculated their average tissue region flux intensities. We found a baseline average standard deviation of 29.9% across all microscopes, scans, and samples, which was reduced to 13.9% after applying sample-specific corrections accounting for differences in the tissue shown on each slide. We used a basic calibration model to correct sample- and microscope-specific effects on overall brightness and relative brightness as a function of the image layer. We tested the generalizability of the calibration procedure and found that applying corrections to independent validation subsets of the samples reduced the variation to 2.9 ± 0.03%. Variations in the unmixed marker expressions were reduced from 15.8% to 4.4% by correcting the raw images to a single reference microscope. Our findings show that mIF microscopes can be standardized for use in clinical pathology laboratories using a relatively simple correction model.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Tipo de estudo: Diagnostic_studies / Prognostic_studies Idioma: En Revista: Cancers (Basel) Ano de publicação: 2023 Tipo de documento: Article País de afiliação: Estados Unidos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Tipo de estudo: Diagnostic_studies / Prognostic_studies Idioma: En Revista: Cancers (Basel) Ano de publicação: 2023 Tipo de documento: Article País de afiliação: Estados Unidos