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Klin Monbl Augenheilkd 2023; 240(12): 1359-1368
DOI: 10.1055/a-2111-8415
Übersicht

Ex Vivo Micro-CT in Ophthalmology: Preparation and Contrasting for Non-invasive 3D-Visualisation

Article in several languages: deutsch | English
Jonas Keiler 
1   Institut für Anatomie, Universitätsmedizin Rostock, Deutschland
,
Thomas Stahnke 
2   Klinik und Poliklinik für Augenheilkunde, Universitätsmedizin Rostock, Deutschland
3   Institut für ImplantatTechnologie und Biomaterialien e. V., Warnemünde, Deutschland
,
Rudolf F. Guthoff
2   Klinik und Poliklinik für Augenheilkunde, Universitätsmedizin Rostock, Deutschland
,
Andreas Wree
1   Institut für Anatomie, Universitätsmedizin Rostock, Deutschland
,
Jens Runge 
1   Institut für Anatomie, Universitätsmedizin Rostock, Deutschland
2   Klinik und Poliklinik für Augenheilkunde, Universitätsmedizin Rostock, Deutschland
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Abstract

X-ray-based micro-computed tomography (micro-CT) is a largely non-destructive imaging method for the visualisation and analysis of internal structures in the ex vivo eye and affords high resolution. In contrast to other high-resolution imaging methods, micro-CT enables spatial recording of larger and more complex tissue structures, such as the anterior chamber of the eye. Special contrasting methods help to enhance the absorption properties of soft tissue, that is otherwise only weakly radiopaque. Critical point drying (CPD), as primarily used in scanning electron microscopy, offers an additional tool for improving differential contrast properties in soft tissue. In the visualisation of intraosseous soft tissue, such as the efferent lacrimal ducts, sample treatment by decalcification with ethylenediaminetetraacetic acid and subsequent CPD provides good results for micro-CT. Micro-CT can be used for a wide range of questions in 1. basic research, 2. application-related studies in ophthalmology (e.g. evaluation of the preclinical application of microstents for glaucoma treatment or analysis of the positioning of intraocular lenses) but also 3. as a supplement to ophthalmological histopathology.

Key words

anterior chamber - iridocorneal angle - lacrimal duct - micro-computed tomography - ocular structures - critical point drying


Publication History

Received: 16 June 2023

Accepted: 06 October 2023

Article published online:
13 December 2023

© 2023. Thieme. All rights reserved.

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

 

  • References/Literatur

  • 1 Gibson E, Gaed M, Gómez JA. et al. 3D prostate histology image reconstruction: Quantifying the impact of tissue deformation and histology section location. J Pathol Inform 2013; 4: 31
    CrossrefPubMedSearch in Google Scholar
  • 2 Hann CR, Bentley MD, Vercnocke A. et al. Imaging the aqueous humor outflow pathway in human eyes by three-dimensional micro-computed tomography (3D micro-CT). Exp Eye Res 2011; 92: 104-111
    CrossrefPubMedSearch in Google Scholar
  • 3 Enders C, Braig EM, Scherer K. et al. Advanced Non-Destructive Ocular Visualization Methods by Improved X-Ray Imaging Techniques. PLoS One 2017; 12: e0170633
    CrossrefPubMedSearch in Google Scholar
  • 4 Leszczyński B, Sojka-Leszczyńska P, Wojtysiak D. et al. Visualization of porcine eye anatomy by X-ray microtomography. Exp Eye Res 2018; 167: 51-55
    CrossrefPubMedSearch in Google Scholar
  • 5 Tkachev SY, Mitrin BI, Karnaukhov NS. et al. Visualization of different anatomical parts of the enucleated human eye using X-ray micro-CT imaging. Exp Eye Res 2021; 203: 108394
    CrossrefPubMedSearch in Google Scholar
  • 6 Runge J, Stahnke T, Guthoff RF. et al. Micro-CT in ophthalmology: ex vivo preparation and contrasting methods for detailed 3D-visualization of eye anatomy with special emphasis on critical point drying. Quant Imaging Med Surg 2022; 12: 4361-4376
    CrossrefPubMedSearch in Google Scholar
  • 7 Hann CR, Vercnocke AJ, Bentley MD. et al. Anatomic changes in Schlemmʼs canal and collector channels in normal and primary open-angle glaucoma eyes using low and high perfusion pressures. Invest Ophthalmol Vis Sci 2014; 55: 5834-5841
    CrossrefPubMedSearch in Google Scholar
  • 8 Lifton JJ, Malcolm AA, McBride JW. An experimental study on the influence of scatter and beam hardening in x-ray CT for dimensional metrology. Meas Sci Technol 2016; 27: 015007
    CrossrefPubMedSearch in Google Scholar
  • 9 Guzman Aparicio MA, Chen TC. New views on three-dimensional imaging technologies for glaucoma: An overview. Curr Opin Ophthalmol 2022; 33: 103-111
    CrossrefPubMedSearch in Google Scholar
  • 10 Bohn S, Stahnke T, Sperlich K. et al. In vivo Histology of the Cornea – From the Rostock Cornea Module to the Rostock Electronic Slit Lamp – A Clinical Proof of Concept Study. Klin Monbl Augenheilkd 2020; 237: 1442-1454
    Thieme ConnectPubMedSearch in Google Scholar
  • 11 Glarin RK, Nguyen BN, Cleary JO. et al. MR-EYE: High-Resolution MRI of the Human Eye and Orbit at Ultrahigh Field (7 T). Magn Reson Imaging Clin N Am 2021; 29: 103-116
    CrossrefPubMedSearch in Google Scholar
  • 12 Helms RW, Minhaz AT, Wilson DL. et al. Clinical 3d imaging of the anterior segment with ultrasound biomicroscopy. Transl Vis Sci Technol 2021; 10: 1-12
    CrossrefPubMedSearch in Google Scholar
  • 13 Yakovlev MA, Vanselow DJ, Ngu MS. et al. A wide-field micro-computed tomography detector: micron resolution at half-centimetre scale. J Synchrotron Radiat 2022; 29: 505-514
    CrossrefPubMedSearch in Google Scholar
  • 14 Aumann S, Donner S, Fischer J. et al. Optical Coherence Tomography (OCT): Principle and Technical Realization. In: High Resolution Imaging in Microscopy and Ophthalmology: New Frontiers in Biomedical Optics [Internet] Cham: Springer International Publishing; 2019: 59-85
    Search in Google Scholar
  • 15 Fouquet C, Gilles JF, Heck N. et al. Improving axial resolution in confocal microscopy with new high refractive index mounting media. PLoS One 2015; 10: 1-17
    CrossrefPubMedSearch in Google Scholar
  • 16 Wang S, Larina IV. High-resolution imaging techniques in tissue engineering. In: Narayan RJ. ed. Monitoring and Evaluation of Biomaterials and their Performance in Vivo. Sawston, Cambridge: Woodhead Publishing; 2017
    CrossrefSearch in Google Scholar
  • 17 Niendorf T, Paul K, Graessl A. et al. Ophthalmologische Bildgebung mit Ultrahochfeld-Magnetresonanztomografie: technische Innovationen und wegweisende Anwendungen. Klin Monbl Augenheilkd 2014; 231: 1187-1195
    Thieme ConnectPubMedSearch in Google Scholar
  • 18 Runge J, Kischkel S, Keiler J. et al. Experimental glaucoma microstent implantation in two animal models and human donor eyes – an ex vivo micro-CT based evaluation of applicability. Quant Imaging Med Surg [accepted].
    PubMed
  • 19 Metscher BD. Micro CT for comparative morphology: Simple staining methods allow high-contrast 3D imaging of diverse non-mineralized animal tissues. BMC Physiol 2009; 9: 11
    CrossrefPubMedSearch in Google Scholar
  • 20 Pauwels E, Van Loo D, Cornillie P. et al. An exploratory study of contrast agents for soft tissue visualization by means of high resolution X-ray computed tomography imaging. J Microsc 2013; 250: 21-31
    CrossrefPubMedSearch in Google Scholar
  • 21 Koç MM, Aslan N, Kao AP. et al. Evaluation of X-ray tomography contrast agents: A review of production, protocols, and biological applications. Microsc Res Tech 2019; 82: 812-848
    CrossrefPubMedSearch in Google Scholar
  • 22 Keiler J, Richter S, Wirkner CS. Evolutionary morphology of the hemolymph vascular system in hermit and king crabs (Crustacea: Decapoda: Anomala). J Morphol 2013; 274: 759-778
    CrossrefPubMedSearch in Google Scholar
  • 23 Nordestgaard BG, Rostgaard J. Critical-point drying versus freeze drying for scanning electron microscopy: a quantitative and qualitative study on isolated hepatocytes. J Microsc 1985; 137: 189-207
    CrossrefPubMedSearch in Google Scholar
  • 24 Horridge GA, Tamm SL. Critical Point Drying for Scanning Electron Microscopic Study of Ciliary Motion. Science 1969; 163: 817-818
    CrossrefPubMedSearch in Google Scholar
  • 25 Jensen OA, Prause JU, Laursen H. Shrinkage in preparatory steps for SEM. A study on rabbit corneal endothelium. Albrecht von Graefes Arch Klin Exp Ophthalmol 1981; 215: 233-242
    CrossrefPubMedSearch in Google Scholar
  • 26 Maser MD, Trimble JJ. Rapid chemical dehydration of biologic samples for scanning electron microscopy using 2,2-dimethoxypropane. J Histochem Cytochem 1977; 25: 247-251
    CrossrefPubMedSearch in Google Scholar
  • 27 Conway K, Kiernan JA. Chemical dehydration of specimens with 2,2-dimethoxypropane (DMP) for paraffin processing of animal tissues: Practical and economic advantages over dehydration in ethanol. Biotech Histochem 1999; 74: 20-26
    CrossrefPubMedSearch in Google Scholar
  • 28 Sun N, Shibata B, Hess JF. et al. An alternative means of retaining ocular structure and improving immunoreactivity for light microscopy studies. Mol Vis 2015; 21: 428-442
    PubMedSearch in Google Scholar
  • 29 Tran H, Jan NJ, Hu D. et al. Formalin Fixation and Cryosectioning Cause only Minimal Changes in Shape or Size of Ocular Tissues. Sci Rep 2017; 7: 1-11
    CrossrefPubMedSearch in Google Scholar
  • 30 Hedrick BP, Yohe L, Vander Linden A. et al. Assessing Soft-Tissue Shrinkage Estimates in Museum Specimens Imaged With Diffusible Iodine-Based Contrast-Enhanced Computed Tomography (diceCT). Microsc Microanal 2018; 24: 284-291
    CrossrefPubMedSearch in Google Scholar
  • 31 Dawood Y, Hagoort J, Siadari BA. et al. Reducing soft-tissue shrinkage artefacts caused by staining with Lugolʼs solution. Sci Rep 2021; 11: 1-13
    CrossrefPubMedSearch in Google Scholar
  • 32 Wree A, Schulze M, Keiler J. Macroscopic anatomy of the nasolacrimal system. In: Anatomische Gesellschaft: 113th Annual Meeting, Rostock 2018. Lecture Abstracts 2018
    Search in Google Scholar
  • 33 Bissinger O, Götz C, Wolff KD. et al. Fully automated segmentation of callus by micro-CT compared to biomechanics. J Orthop Surg Res 2017; 12: 1-9
    CrossrefPubMedSearch in Google Scholar
  • 34 Ferl GZ, Barck KH, Patil J. et al. Automated segmentation of lungs and lung tumors in mouse micro-CT scans. iScience 2022; 25: 105712
    CrossrefPubMedSearch in Google Scholar
  • 35 Vincenzi E, Fantazzini A, Basso C. et al. A fully automated deep learning pipeline for micro-CT-imaging-based densitometry of lung fibrosis murine models. Respir Res 2022; 23: 1-14
    CrossrefPubMedSearch in Google Scholar
  • 36 Malimban J, Lathouwers D, Qian H. et al. Deep learning-based segmentation of the thorax in mouse micro-CT scans. Sci Rep 2022; 12: 1-12
    CrossrefPubMedSearch in Google Scholar
  • 37 Barillot C, Gibaud B, Lis O. et al. Computer graphics in medicine: a survey. Crit Rev Biomed Eng 1988; 15: 269-307
    PubMedSearch in Google Scholar
  • 38 Palladino A, Salerno A, Crasto A. et al. Integration of micro-CT and histology data for vasculature morpho-functional analysis in tissue regeneration. Ann Anat 2023; 245: 152019
    CrossrefPubMedSearch in Google Scholar
  • 39 Chicherova N, Hieber SE, Khimchenko A. et al. Automatic deformable registration of histological slides to µCT volume data. J Microsc 2018; 271: 49-61
    CrossrefPubMedSearch in Google Scholar
  • 40 Lupariello F, Genova T, Mussano F. et al. Micro-CT processingʼs effects on microscopic appearance of human fetal cardiac samples. Leg Med 2021; 53: 101934
    CrossrefPubMedSearch in Google Scholar