ArticlePloS one2012
A novel immunofluorescent computed tomography (ICT) method to localise and quantify multiple antigens in large tissue volumes at high resolution.
Article in PloS one, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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Who cites it
25 citing papers in PubMed.
- Isolation and Culture of Human Meibomian Gland Ductal Cells.Investigative ophthalmology & visual science · 2023Article
- Meibomian gland stem/progenitor cells: The hunt for gland renewal.The ocular surface · 2023Article
- Immuno Tomography (IT) and Imaging Mass Cytometry (IMC) for constructing spatially resolved, multiplexed 3D IMC data sets.The ocular surface · 2022Article
- Characterisation of carapace composition in developing and adult ostracods (Marine biology · 2022Article
- Hyperkeratinization and Proinflammatory Cytokine Expression in Meibomian Glands Induced by Staphylococcus aureus.Investigative ophthalmology & visual science · 2021Article
- EPHA2-dependent outcompetition of KRASG12D mutant cells by wild-type neighbors in the adult pancreas.Current biology : CB · 2021Article
- Novel approach for quantification of multiple immunofluorescent signals using histograms and 2D plot profiling of whole-section panoramic images.Scientific reports · 2021Article
- Meibomian Gland Dysfunction: What Have Animal Models Taught Us?International journal of molecular sciences · 2020Review
- Organotypic Culture of Mouse Meibomian Gland: A Novel Model to Study Meibomian Gland Dysfunction In Vitro.Investigative ophthalmology & visual science · 2020Article
- Epithelial Migration and Non-adhesive Periderm Are Required for Digit Separation during Mammalian Development.Developmental cell · 2020Article
- Immunofluorescence Tomography: High-resolution 3-D reconstruction by serial-sectioning of methacrylate embedded tissues and alignment of 2-D immunofluorescence images.Scientific reports · 2019Article
- Proteolytic activity in the meibomian gland: Implications to health and disease.Experimental eye research · 2017Review
- Meibocyte differentiation and renewal: Insights into novel mechanisms of meibomian gland dysfunction (MGD).Experimental eye research · 2017Review
- Ocular surface inflammation impairs structure and function of meibomian gland.Experimental eye research · 2017Review
- A Tool for Brain-Wide Quantitative Analysis of Molecular Data upon Projection into a Planar View of Choice.Frontiers in neuroanatomy · 2017Article
- Renewal of the Holocrine Meibomian Glands by Label-Retaining, Unipotent Epithelial Progenitors.Stem cell reports · 2016Article
- Conventional fluorescence microscopy below the diffraction limit with simultaneous capture of two fluorophores in DNA origami.Proceedings of SPIE--the International Society for Optical Engineering · 2016Article
- Simultaneous two color image capture for sub-diffraction localization fluorescence microscopy.Micron (Oxford, England : 1993) · 2016Article
- Meibomian gland dysfunction: hyperkeratinization or atrophy?BMC ophthalmology · 2015Review
- Immunofluorescence Tomography of Mouse Ocular Surface Epithelial Stem Cells and Their Niche Microenvironment.Investigative ophthalmology & visual science · 2015Article
Corrections and comments
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Authors and funding
6 authors.
Funding
Abstract
Current immunofluorescence protocols are limited as they do not provide reliable antibody staining within large tissue volumes (mm(3)) and cannot localise and quantify multiple antigens or cell populations in the same tissue at high resolution. To address this limitation, we have developed an approach to three-dimensionally visualise large tissue volumes (mm(3)) at high resolution (<1 µm) and with multiple antigen labelling, for volumetric and quantitative analysis. This is made possible through computer reconstruction of serial sectioned and sequentially immunostained butyl-methyl methacrylate (BMMA) embedded tissue. Using this novel immunofluorescent computed tomography (ICT) approach, we have three-dimensionally reconstructed part of the murine lower eyelid that contains the meibomian gland and localised cell nuclei (DAPI), Ki67 and cytokeratin 1 (CK1), as well as performing non-linear optical (NLO) microscopy imaging of collagen, to assess cell density, cell proliferation, gland keratinisation and gland volume respectively. Antigenicity was maintained after four iterative stains on the same tissue, suggesting that there is no defined limit to the number of antigens that can be immunostained for reconstruction, as long as the sections remain intact and the previous antibody has been successfully eluted. BMMA resin embedding also preserved fluorescence of transgenic proteins. We propose that ICT may provide valuable high resolution, three-dimensional biological maps of multiple biomolecules within a single tissue or organ to better characterise and quantify tissue structure and function.
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