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Anatomia, histologia, embryologia2022; 51(5); 611-623; doi: 10.1111/ahe.12834

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

Abstract: 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.
© 2022 The Authors. Anatomia, Histologia, Embryologia published by Wiley-VCH GmbH.
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Publication Date: 2022-07-18 PubMed ID: 35851500PubMed Central: PMC9542120DOI: 10.1111/ahe.12834Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The researchers have developed an optimized workflow for microCT imaging of formalin-fixed and paraffin-embedded (FFPE) early horse embryos. The method is designed to provide high-detailed imaging while maintaining the integrity of the embryos for subsequent analyses.

Research Background and Objective

  • This study presents a method to improve the imaging of horse embryos fixed in formalin and embedded in paraffin, focusing primarily on embryos gathered on the 34th day of pregnancy—a crucial point just before the formation of the placenta.
  • The imaging technique is also designed for use with larger animals’ embryos or generally, any FFPE tissue specimens to examine their morphology and developmental aspects.
  • Microscopic 3D imaging techniques such as microCT are essential in studying normal embryogenesis and developmental disorders.

Improved Imaging Procedure

  • Unlike previous microCT imaging attempts on vertebrate embryos which required staining with an X-ray dense contrast agent, the technique presented here allows visualizing the morphology and organ development in unstained FFPE embryos.
  • The team paid particular attention to two considerations for high-quality data acquisition—minimizing paraffin around the sample and improving the signal-to-noise ratio through image filtering, as FFPE embryos generally produce low-contrast data sets.

Workflow and Applicability

  • The researchers developed and implemented a workflow that aids in capturing a comprehensive imaging of the embryo and assists in analyzing organ volumes and development.
  • High-resolution interior tomographies obtained using this method can also provide valuable ‘virtual histology’ information from selected regions of interest.
  • Another significant advantage of this imaging technique is that the embryos remain intact during the scanning process, thus, enabling them for additional studies using conventional histology or immunohistochemistry methods.
  • This characteristic also allows application of the workflow to archival paraffin-embedded material, hence increasing its versatility and potential uses in research.

Cite This Article

APA
Handschuh S, Okada CTC, Walter I, Aurich C, Glösmann M. (2022). An optimized workflow for microCT imaging of formalin-fixed and paraffin-embedded (FFPE) early equine embryos. Anat Histol Embryol, 51(5), 611-623. https://doi.org/10.1111/ahe.12834

Publication

Anatomia, histologia, embryologia
ISSN: 1439-0264
NlmUniqueID: 7704218
Country: Germany
Language: English
Volume: 51
Issue: 5
Pages: 611-623

Researcher Affiliations

Handschuh, Stephan
  • VetCore Facility for Research/Imaging Unit, University of Veterinary Medicine Vienna, Vienna, Austria.
Okada, Carolina T C
  • Platform Artificial Insemination and Embryo Transfer, Department for Small Animals and Horses, University of Veterinary Medicine Vienna, Vienna, Austria.
Walter, Ingrid
  • VetCore Facility for Research/VetBiobank, University of Veterinary Medicine Vienna, Vienna, Austria.
  • Institute of Morphology, University of Veterinary Medicine Vienna, Vienna, Austria.
Aurich, Christine
  • Platform Artificial Insemination and Embryo Transfer, Department for Small Animals and Horses, University of Veterinary Medicine Vienna, Vienna, Austria.
Glösmann, Martin
  • VetCore Facility for Research/Imaging Unit, University of Veterinary Medicine Vienna, Vienna, Austria.

MeSH Terms

  • Animals
  • Formaldehyde
  • Horses
  • Paraffin Embedding / veterinary
  • Tissue Fixation / veterinary
  • Vertebrates
  • Workflow
  • X-Ray Microtomography / methods
  • X-Ray Microtomography / veterinary

Grant Funding

  • University of Veterinary Medicine Vienna

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Citations

This article has been cited 5 times.
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