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An optimized workflow for microCT imaging of formalin‐fixed and paraffin‐embedded (FFPE) early equine embryos

Here, we describe a workflow for high‐detail microCT imaging of formalin‐fixed and paraffin‐embedded (FFPE) equine embryos recovered on Day 34 of pregnancy (E34), a period just before placenta formation. The presented imaging methods are suitable for large animals' embryos with intention to stu...

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Detalles Bibliográficos
Autores principales: Handschuh, Stephan, Okada, Carolina T. C., Walter, Ingrid, Aurich, Christine, Glösmann, Martin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9542120/
https://www.ncbi.nlm.nih.gov/pubmed/35851500
http://dx.doi.org/10.1111/ahe.12834
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author Handschuh, Stephan
Okada, Carolina T. C.
Walter, Ingrid
Aurich, Christine
Glösmann, Martin
author_facet Handschuh, Stephan
Okada, Carolina T. C.
Walter, Ingrid
Aurich, Christine
Glösmann, Martin
author_sort Handschuh, Stephan
collection PubMed
description Here, we describe a workflow for high‐detail microCT imaging of formalin‐fixed and paraffin‐embedded (FFPE) equine embryos recovered on Day 34 of pregnancy (E34), a period just before placenta formation. The presented imaging methods are suitable for large animals' embryos with intention to study morphological and developmental aspects, but more generally can be adopted for all kinds of FFPE tissue specimens. Microscopic 3D imaging techniques such as microCT are important tools for detecting and studying normal embryogenesis and developmental disorders. To date, microCT imaging of vertebrate embryos was mostly done on embryos that have been stained with an X‐ray dense contrast agent. Here, we describe an alternative imaging procedure that allows to visualize embryo morphology and organ development in unstained FFPE embryos. Two aspects are critical for high‐quality data acquisition: (i) a proper sample mounting leaving as little as possible paraffin around the sample and (ii) an image filtering pipeline that improves signal‐to‐noise ratio in these inherently low‐contrast data sets. The presented workflow allows overview imaging of the whole embryo proper and can be used for determination of organ volumes and development. Furthermore, we show that high‐resolution interior tomographies can provide virtual histology information from selected regions of interest. In addition, we demonstrate that microCT scanned embryos remain intact during the scanning procedure allowing for a subsequent investigation by routine histology and/or immunohistochemistry. This makes the presented workflow applicable also to archival paraffin‐embedded material.
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spelling pubmed-95421202022-10-14 An optimized workflow for microCT imaging of formalin‐fixed and paraffin‐embedded (FFPE) early equine embryos Handschuh, Stephan Okada, Carolina T. C. Walter, Ingrid Aurich, Christine Glösmann, Martin Anat Histol Embryol Original Articles Here, we describe a workflow for high‐detail microCT imaging of formalin‐fixed and paraffin‐embedded (FFPE) equine embryos recovered on Day 34 of pregnancy (E34), a period just before placenta formation. The presented imaging methods are suitable for large animals' embryos with intention to study morphological and developmental aspects, but more generally can be adopted for all kinds of FFPE tissue specimens. Microscopic 3D imaging techniques such as microCT are important tools for detecting and studying normal embryogenesis and developmental disorders. To date, microCT imaging of vertebrate embryos was mostly done on embryos that have been stained with an X‐ray dense contrast agent. Here, we describe an alternative imaging procedure that allows to visualize embryo morphology and organ development in unstained FFPE embryos. Two aspects are critical for high‐quality data acquisition: (i) a proper sample mounting leaving as little as possible paraffin around the sample and (ii) an image filtering pipeline that improves signal‐to‐noise ratio in these inherently low‐contrast data sets. The presented workflow allows overview imaging of the whole embryo proper and can be used for determination of organ volumes and development. Furthermore, we show that high‐resolution interior tomographies can provide virtual histology information from selected regions of interest. In addition, we demonstrate that microCT scanned embryos remain intact during the scanning procedure allowing for a subsequent investigation by routine histology and/or immunohistochemistry. This makes the presented workflow applicable also to archival paraffin‐embedded material. John Wiley and Sons Inc. 2022-07-18 2022-09 /pmc/articles/PMC9542120/ /pubmed/35851500 http://dx.doi.org/10.1111/ahe.12834 Text en © 2022 The Authors. Anatomia, Histologia, Embryologia published by Wiley‐VCH GmbH. https://creativecommons.org/licenses/by-nc/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ (https://creativecommons.org/licenses/by-nc/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.
spellingShingle Original Articles
Handschuh, Stephan
Okada, Carolina T. C.
Walter, Ingrid
Aurich, Christine
Glösmann, Martin
An optimized workflow for microCT imaging of formalin‐fixed and paraffin‐embedded (FFPE) early equine embryos
title An optimized workflow for microCT imaging of formalin‐fixed and paraffin‐embedded (FFPE) early equine embryos
title_full An optimized workflow for microCT imaging of formalin‐fixed and paraffin‐embedded (FFPE) early equine embryos
title_fullStr An optimized workflow for microCT imaging of formalin‐fixed and paraffin‐embedded (FFPE) early equine embryos
title_full_unstemmed An optimized workflow for microCT imaging of formalin‐fixed and paraffin‐embedded (FFPE) early equine embryos
title_short An optimized workflow for microCT imaging of formalin‐fixed and paraffin‐embedded (FFPE) early equine embryos
title_sort optimized workflow for microct imaging of formalin‐fixed and paraffin‐embedded (ffpe) early equine embryos
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9542120/
https://www.ncbi.nlm.nih.gov/pubmed/35851500
http://dx.doi.org/10.1111/ahe.12834
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