RSS-Feed abonnieren
DOI: 10.1055/a-2111-8415
Ex-vivo-Mikro-CT in der Augenheilkunde: Probenbehandlung und -kontrastierung für die 3D-Darstellung
Artikel in mehreren Sprachen: deutsch | English![](https://www.thieme-connect.de/media/klimo/202312/lookinside/thumbnails/10-1055-a-2111-8415_kl2884_de-1.jpg)
Zusammenfassung
Die röntgenbasierte Mikrocomputertomografie (Mikro-CT) bietet ein weitgehend zerstörungsfreies Bildgebungsverfahren zur Visualisierung und Analyse interner Strukturen im Ex-vivo-Auge mit hohen Auflösungen. Im Gegensatz zu anderen hochauflösenden Imaging-Verfahren ermöglicht die Mikro-CT die räumliche Erfassung größerer und komplexerer Gewebestrukturen wie der vorderen Augenkammer. Spezielle Kontrastierungsmethoden helfen, die Absorptionseigenschaften von ansonsten nur schwach röntgenopaken Weichteilgeweben zu verstärken. Die vor allem in der Rasterelektronenmikroskopie verwendete Kritisch-Punkt-Trocknung (KPT) bietet ein zusätzliches Werkzeug zur Verbesserung der differenziellen Kontrasteigenschaften im Weichteilgewebe. Bei der Darstellung intraossärer Weichteilgewebe, wie den ableitenden Tränenwegen, liefert die Probenbehandlung durch Entkalkung mit Ethylendiamintetraessigsäure und anschließender KPT gute Ergebnisse für die Mikro-CT. Die Mikro-CT kann für verschiedenste Fragestellungen in 1. Grundlagenforschung, 2. anwendungsbezogenen Studien in der Augenheilkunde (z. B. Evaluierung der präklinischen Applikation von Microstents zur Glaukombehandlung oder Analyse der Positionierung von Intraokularlinsen), aber auch 3. als Ergänzung für die ophthalmologische Histopathologie eingesetzt werden.
Schlüsselwörter
vordere Augenkammer - ableitende Tränenwege - Mikrocomputertomografie - okulare Strukturen - Kammerwinkel - Kritisch-Punkt-TrocknungPublikationsverlauf
Eingereicht: 16. Juni 2023
Angenommen: 06. Oktober 2023
Artikel online veröffentlicht:
13. Dezember 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 DOI: 10.4103/2153-3539.120874.
- 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 DOI: 10.1016/j.exer.2010.12.010.
- 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 DOI: 10.1371/journal.pone.0170633.
- 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 DOI: 10.1016/j.exer.2017.11.004.
- 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 DOI: 10.1016/j.exer.2020.108394.
- 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 DOI: 10.21037/qims-22-109.
- 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 DOI: 10.1167/iovs.14-14128.
- 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 DOI: 10.1088/0957-0233/27/1/015007.
- 9 Guzman Aparicio MA, Chen TC. New views on three-dimensional imaging technologies for glaucoma: An overview. Curr Opin Ophthalmol 2022; 33: 103-111 DOI: 10.1097/ICU.0000000000000828.
- 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 DOI: 10.1055/a-1297-4717.
- 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 DOI: 10.1016/j.mric.2020.09.004.
- 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 DOI: 10.1167/tvst.10.3.11.
- 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 DOI: 10.1107/S160057752101287X.
- 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
- 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 DOI: 10.1371/journal.pone.0121096.
- 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
- 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 DOI: 10.1055/s-0034-1383365.
- 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].
- 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 DOI: 10.1186/1472-6793-9-11.
- 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 DOI: 10.1111/jmi.12013.
- 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 DOI: 10.1002/jemt.23225.
- 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
- 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 DOI: 10.1111/j.1365-2818.1985.tb02577.x.
- 24 Horridge GA, Tamm SL. Critical Point Drying for Scanning Electron Microscopic Study of Ciliary Motion. Science 1969; 163: 817-818 DOI: 10.1126/science.163.3869.817.
- 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 DOI: 10.1007/BF00407662.
- 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 DOI: 10.1177/25.4.67137.
- 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 DOI: 10.3109/10520299909066473.
- 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
- 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 DOI: 10.1038/s41598-017-12006-1.
- 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 DOI: 10.1017/S1431927618000399.
- 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 DOI: 10.1038/s41598-021-99202-2.
- 32 Wree A, Schulze M, Keiler J. Macroscopic anatomy of the nasolacrimal system. In: Anatomische Gesellschaft: 113th Annual Meeting, Rostock 2018. Lecture Abstracts 2018
- 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 DOI: 10.1186/s13018-017-0609-9.
- 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 DOI: 10.1016/j.isci.2022.105712.
- 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 DOI: 10.1186/s12931-022-02236-x.
- 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 DOI: 10.1038/s41598-022-05868-7.
- 37 Barillot C, Gibaud B, Lis O. et al. Computer graphics in medicine: a survey. Crit Rev Biomed Eng 1988; 15: 269-307
- 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 DOI: 10.1016/j.aanat.2022.152019.
- 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 DOI: 10.1111/jmi.12692.
- 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 DOI: 10.1016/j.legalmed.2021.101934.