Combination of µCT and light microscopy for generation-specific stereological analysis of pulmonary arterial branches: a proof-of-concept study.
3D reconstruction
Branching generation analysis
Light microscopy
Micro computed tomography
Pulmonary vasculature
Stereology
Journal
Histochemistry and cell biology
ISSN: 1432-119X
Titre abrégé: Histochem Cell Biol
Pays: Germany
ID NLM: 9506663
Informations de publication
Date de publication:
Feb 2021
Feb 2021
Historique:
accepted:
12
11
2020
pubmed:
3
12
2020
medline:
3
7
2021
entrez:
2
12
2020
Statut:
ppublish
Résumé
Various lung diseases, including pulmonary hypertension, chronic obstructive pulmonary disease or bronchopulmonary dysplasia, are associated with structural and architectural alterations of the pulmonary vasculature. The light microscopic (LM) analysis of the blood vessels is limited by the fact that it is impossible to identify which generation of the arterial tree an arterial profile within a LM microscopic section belongs to. Therefore, we established a workflow that allows for the generation-specific quantitative (stereological) analysis of pulmonary blood vessels. A whole left rabbit lung was fixed by vascular perfusion, embedded in glycol methacrylate and imaged by micro-computed tomography (µCT). The lung was then exhaustively sectioned and 20 consecutive sections were collected every 100 µm to obtain a systematic uniform random sample of the whole lung. The digital processing involved segmentation of the arterial tree, generation analysis, registration of LM sections with the µCT data as well as registration of the segmentation and the LM images. The present study demonstrates that it is feasible to identify arterial profiles according to their generation based on a generation-specific color code. Stereological analysis for the first three arterial generations of the monopodial branching of the vasculature included volume fraction, total volume, lumen-to-wall ratio and wall thickness for each arterial generation. In conclusion, the correlative image analysis of µCT and LM-based datasets is an innovative method to assess the pulmonary vasculature quantitatively.
Identifiants
pubmed: 33263790
doi: 10.1007/s00418-020-01946-x
pii: 10.1007/s00418-020-01946-x
pmc: PMC7709482
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
227-239Subventions
Organisme : Deutsche Forschungsgemeinschaft
ID : MU3118/8-1
Références
Ackermann M, Stark H, Neubert L, Schubert S, Borchert P, Linz F, Wagner WL, Stiller W, Wielpütz M, Hoefer A, Haverich A, Mentzer SJ, Shah HR, Welte T, Kuehnel M, Jonigk D (2020) Morphomolecular motifs of pulmonary neoangiogenesis in interstitial lung diseases. Eur Respir J 55:1900933. https://doi.org/10.1183/13993003.00933-2019
doi: 10.1183/13993003.00933-2019
pubmed: 31806721
Ackermann M, Verleden SE, Kuehnel M, Haverich A, Welte T, Laenger F, Vanstapel A, Werlein C, Stark H, Tzankov A, Li WW, Li VW, Mentzer SJ, Jonigk D (2020) Pulmonary Vascular Endothelialitis, Thrombosis, and Angiogenesis in Covid-19. N Engl J Med 383:120–128. https://doi.org/10.1056/NEJMoa2015432
doi: 10.1056/NEJMoa2015432
pubmed: 32437596
pmcid: 7412750
Ayachit U (2016) The ParaView guide. Kitware Inc., New York
Baddeley AJ, Gundersen HJ, Cruz-Orive LM (1986) Estimation of surface area from vertical sections. J Microsc 142:259–276. https://doi.org/10.1111/j.1365-2818.1986.tb04282.x
doi: 10.1111/j.1365-2818.1986.tb04282.x
pubmed: 3735415
Brandenberger C, Ochs M, Mühlfeld C (2015) Assessing particle and fiber toxicology in the respiratory system: the stereology toolbox. Part Fibre Toxicol 12:35. https://doi.org/10.1186/s12989-015-0110-8
doi: 10.1186/s12989-015-0110-8
pubmed: 26521139
pmcid: 4628359
Buchacker T, Mühlfeld C, Wrede C, Wagner WL, Beare R, McCormick M, Grothausmann R (2019) Assessment of the Alveolar Capillary Network in the Postnatal Mouse Lung in 3D Using Serial Block-Face Scanning Electron Microscopy. Front Physiol 10:1357. https://doi.org/10.3389/fphys.2019.01357
doi: 10.3389/fphys.2019.01357
pubmed: 31824323
pmcid: 6881265
Davies P, Reid L (1991) Hypoxic remodeling of the rat pulmonary arterial microcirculation assessed by microdissection. J Appl Physiol (1985) 71:1886–1891. https://doi.org/10.1152/jappl.1991.71.5.1886
doi: 10.1152/jappl.1991.71.5.1886
deMello DE, Sawyer D, Galvin N, Reid LM (1997) Early fetal development of lung vasculature. Am J Respir Cell Mol Biol 16:568–581. https://doi.org/10.1165/ajrcmb.16.5.9160839
doi: 10.1165/ajrcmb.16.5.9160839
pubmed: 9160839
Faight EM, Verdelis K, Zourelias L, Chong R, Benza RL, Shields KJ (2017) MicroCT analysis of vascular morphometry: a comparison of right lung lobes in the SUGEN/hypoxic rat model of pulmonary arterial hypertension. Pulm Circ 7:522–530. https://doi.org/10.1177/2045893217709001
doi: 10.1177/2045893217709001
pubmed: 28597764
pmcid: 5467946
Gehr P, Geiser M, Stone KC, Crapo JD (1993) Morphometric analysis of the gas exchange region of the lung. In: Gardner DE, Crapo JD, McClellan RO (eds) Toxicology of the lung, 2nd edn. Raven Press, New York, pp 111–154
Gil J (1990) Controlled and reproducible fixation of the lung for correlated studies. Gil J Models of lung diseases. Microscopy and structural methods, vol 47. Marcel Dekker, New York, pp 3–22
Grothausmann R, Knudsen L, Ochs M, Mühlfeld C (2017) Digital 3D reconstructions using histological serial sections of lung tissue including the alveolar capillary network. Am J Physiol Lung Cell Mol Physiol 312:L243–L257. https://doi.org/10.1152/ajplung.00326.2016
doi: 10.1152/ajplung.00326.2016
pubmed: 27913424
Grothausmann R, Zukić D, McCormick M, Mühlfeld C, Knudsen L (2020) Enabling manual intervention for otherwise automated registration of large image series, biomedical image registration. Springer International Publishing, Berlin, pp 23–33. https://doi.org/10.1007/978-3-030-50120-4_3
Gundersen HJ, Jensen EB (1987) The efficiency of systematic sampling in stereology and its prediction. J Microsc 147:229–263. https://doi.org/10.1111/j.1365-2818.1987.tb02837.x
doi: 10.1111/j.1365-2818.1987.tb02837.x
pubmed: 3430576
Haberthür D, Barré SF, Tschanz SA, Yao E, Stampanoni M, Schittny JC (2013) Visualization and stereological characterization of individual rat lung acini by high-resolution X-ray tomographic microscopy. J Appl Physiol (1985) 115:1379–1387. https://doi.org/10.1152/japplphysiol.00642.2013
doi: 10.1152/japplphysiol.00642.2013
Hlushchuk R, Zubler C, Barré S, Correa Shokiche C, Schaad L, Röthlisberger R, Wnuk M, Daniel C, Khoma O, Tschanz SA, Reyes M, Djonov V (2018) Cutting-edge microangio-CT: new dimensions in vascular imaging and kidney morphometry. Am J Physiol Renal Physiol 314:F493–F499. https://doi.org/10.1152/ajprenal.00099.2017
doi: 10.1152/ajprenal.00099.2017
pubmed: 29167169
Horsfield K (1978) Morphometry of the small pulmonary arteries in man. Circ Res 42:593–597. https://doi.org/10.1161/01.RES.42.5.593
doi: 10.1161/01.RES.42.5.593
pubmed: 639181
Horsfield K (1984) Axial pathways compared with complete data in morphological studies of the lung. Respir Physiol 55:317–324. https://doi.org/10.1016/0034-5687(84)90054-9
doi: 10.1016/0034-5687(84)90054-9
pubmed: 6739988
Hsia CC, Hyde DM, Ochs M, Weibel ER; ATS/ERS Joint Task Force on Quantitative Assessment of Lung Structure (2010) An official research policy statement of the American Thoracic Society/European Respiratory Society: standards for quantitative assessment of lung structure. Am J Respir Crit Care Med 181:394–418. https://doi.org/10.1164/rccm.200809-1522ST
doi: 10.1164/rccm.200809-1522ST
Ibanez L, Lorensen B, McCormick M, King B, Blezek D, Johnson H et al (2020) InsightSoftwareConsortium/ITK: ITK 5.1.0 (Version v5.1.0). Zenodo. https://doi.org/10.5281/zenodo.3888793
doi: 10.5281/zenodo.3888793
Jiménez J, Richter J, Nagatomo T, Salaets T, Quarck R, Wagennar A, Wang H, Vanoirbeek J, Deprest J, Toelen J (2016) Progressive vascular functional and structural damage in a bronchopulmonary dysplasia model in preterm rabbits exposed to hyperoxia. Int J Mol Sci 17:1776. https://doi.org/10.3390/ijms17101776
doi: 10.3390/ijms17101776
Kasahara Y, Tuder RM, Cool CD, Lynch DA, Flores SC, Voelkel NF (2001) Endothelial cell death and decreased expression of vascular endothelial growth factor and vascular endothelial growth factor receptor 2 in emphysema. Am J Respir Crit Care Med 163:737–744. https://doi.org/10.1164/ajrccm.163.3.2002117
doi: 10.1164/ajrccm.163.3.2002117
pubmed: 11254533
Lee D, Willits N, Wexler AS (2011) Detecting alterations in pulmonary airway development with airway-by-airway comparison. Ann Biomed Eng 39:1805–1814. https://doi.org/10.1007/s10439-011-0279-4
doi: 10.1007/s10439-011-0279-4
pubmed: 21347548
pmcid: 3096773
Madl P, Hofmann W, Oldham MJ, Asgharian B (2010) Stochastic morphometric model of the BALB/c mouse lung. Anat Rec (Hoboken) 293:1766–1775. https://doi.org/10.1002/ar.21208
doi: 10.1002/ar.21208
Marstal K, Berendsen F, Staring M, Klein S (2016) SimpleElastix: A user-friendly, multi-lingual library for medical image registration. In: International Workshop on Biomedical Image Registration (WBIR), pp 574582, https://doi.org/10.1109/CVPRW.2016.78
Mayhew TM (2008) Taking tissue samples from the placenta: an illustration of principles and strategies. Placenta 29:1–14. https://doi.org/10.1016/j.placenta.2007.05.010
doi: 10.1016/j.placenta.2007.05.010
pubmed: 17658596
Mühlfeld C, Ochs M (2013) Quantitative microscopy of the lung: a problem-based approach. Part 2: stereological parameters and study designs in various diseases of the respiratory tract. Am J Physiol Lung Cell Mol Physiol 305:L205-221
doi: 10.1152/ajplung.00427.2012
Mühlfeld C, Wrede C, Knudsen L, Buchacker T, Ochs M, Grothausmann R (2018) Recent developments in 3-D reconstruction and stereology to study the pulmonary vasculature. Am J Physiol Lung Cell Mol Physiol 315:L173–L183
doi: 10.1152/ajplung.00541.2017
Ochs M, Weibel ER (2008) Functional design of the human lung for gas exchange. In: Fishman AP, Elias JA, Fishman JA, Grippi MA, Senior RM, Pack AI (eds) Fishman’s pulmonary diseases and disorders, 4th edn. McGraw-Hill, New York, pp 23–69
Ochs M, Mühlfeld C (2013) Quantitative microscopy of the lung—a problem-based approach. Part 1: basic principles of stereology. Am J Physiol Lung Cell Mol Physiol 305:L15–L22. https://doi.org/10.1152/ajplung.00429.2012
doi: 10.1152/ajplung.00429.2012
pubmed: 23624789
Saccomano M, Albers J, Tromba G, Dobrivojević Radmilović M, Gajović S, Alves F, Dullin C (2018) Synchrotron inline phase contrast µCT enables detailed virtual histology of embedded soft-tissue samples with and without staining. J Synchrotron Radiat 25:1153–1161. https://doi.org/10.1107/S1600577518005489
doi: 10.1107/S1600577518005489
pubmed: 29979177
Schneider JP, Ochs M (2014) Alterations of mouse lung tissue dimensions during processing for morphometry: a comparison of methods. Am J Physiol Lung Cell Mol Physiol 306:L341-350. https://doi.org/10.1152/ajplung.00329.2013
doi: 10.1152/ajplung.00329.2013
pubmed: 24375800
Schroeder W, Martin K, Lorensen B (2006) The visualization toolkit: an object-oriented approach to 3D graphics. Kitware Inc., New York
Singhal S, Henderson R, Horsfield K, Harding K (1973) Cumming G (1973) Morphometry of the human pulmonary arterial tree. Circ Res 33:190–197. https://doi.org/10.1161/01.res.33.2.190
doi: 10.1161/01.res.33.2.190
pubmed: 4727370
Tange O (2011) GNU Parallel—the command-line power tool; login. USENIX Mag 36:42–47
Thébaud B, Abman SH (2007) Bronchopulmonary dysplasia: where have all the vessels gone? Roles of angiogenic growth factors in chronic lung disease. Am J Respir Crit Care Med 175:978–985. https://doi.org/10.1164/rccm.200611-1660PP
doi: 10.1164/rccm.200611-1660PP
pubmed: 17272782
pmcid: 2176086
Townsley MI (2012) Structure and composition of pulmonary arteries, capillaries, and veins. Compr Physiol 2:675–709. https://doi.org/10.1002/cphy.c100081
doi: 10.1002/cphy.c100081
pubmed: 23606929
pmcid: 3630377
Tuder RM (2017) Pulmonary vascular remodeling in pulmonary hypertension. Cell Tissue Res 367:643–649. https://doi.org/10.1007/s00441-016-2539-y
doi: 10.1007/s00441-016-2539-y
pubmed: 28025704
Vasilescu DM, Gao Z, Saha PK, Yin L, Wang G, Haefeli-Bleuer B, Ochs M, Weibel ER, Hoffman EA (2012) Assessment of morphometry of pulmonary acini in mouse lungs by nondestructive imaging using multi- scale microcomputed tomography. Proc Natl Acad SciUSA 109:17105–17110. https://doi.org/10.1073/pnas.1215112109
doi: 10.1073/pnas.1215112109
Vasilescu DM, Klinge C, Knudsen L, Yin L, Wang G, Weibel ER, Ochs M, Hoffman EA (2013) Stereological assessment of mouse lung parenchyma via nondestructive, multiscale micro-CT imaging validated by light microscopic histology. J Appl Physiol 114:716–724. https://doi.org/10.1152/japplphysiol.00855.2012
doi: 10.1152/japplphysiol.00855.2012
pubmed: 23264542
Wang PM, Kraman SS (2004) Fractal branching pattern of the monopodial canine airway. J Appl Physiol (1985) 96:2194–2199. https://doi.org/10.1152/japplphysiol.00604.2003
doi: 10.1152/japplphysiol.00604.2003
Weibel ER (1984) Morphometric and stereological methods in respiratory physiology, including fixation techniques. In: Otis AB (ed) Techniques in the life sciences: respiratory physiology. Elsevier, County Clare, pp 1–35
Yushkevich PA, Piven J, Hazlett HC, Smith RG, Ho S, Gee JC, Gerig G (2006) User-guided 3D active contour segmentation of anatomical structures: significantly improved efficiency and reliability. Neuroimage 31:1116–1128. https://doi.org/10.1016/j.neuroimage.2006.01.015
doi: 10.1016/j.neuroimage.2006.01.015
pubmed: 16545965