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Article

3D Visualization of Human Blood Vascular Networks Using Single-Domain Antibodies Directed against Endothelial Cell-Selective Adhesion Molecule (ESAM)

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Research Group ‘Lymphovascular Medicine and Translational 3D-Histopathology’, Institute of Medical and Human Genetics, Charité, Universitätsmedizin Berlin, Augustenburger Platz 1, 13353 Berlin, Germany
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Berlin Institute of Health at Charité-Universitätsmedizin Berlin, BIH Center for Regenerative Therapies, Augustenburger Platz 1, 13353 Berlin, Germany
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Research Group ‘Development and Disease’, Max Planck Institute for Molecular Genetics, Ihnestraße 63-73, 14195 Berlin, Germany
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Department of Rheumatology and Clinical Immunology, Maasstad Hospital, Maasstadweg 21, 3079 DZ Rotterdam, The Netherlands
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Department of Immunology, Erasmus University Medical Center, Doctor Molewaterplein 40, 3015 GD Rotterdam, The Netherlands
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Berlin Institute of Health at Charité, Universitätsmedizin Berlin, BIH Academy, Clinician Scientist Program, Charitéplatz 1, 10117 Berlin, Germany
*
Author to whom correspondence should be addressed.
Academic Editor: Daniel Henrion
Int. J. Mol. Sci. 2022, 23(8), 4369; https://doi.org/10.3390/ijms23084369
Received: 18 March 2022 / Revised: 7 April 2022 / Accepted: 12 April 2022 / Published: 15 April 2022
(This article belongs to the Section Molecular Biology)
High-quality three-dimensional (3D) microscopy allows detailed, unrestricted and non-destructive imaging of entire volumetric tissue specimens and can therefore increase the diagnostic accuracy of histopathological tissue analysis. However, commonly used IgG antibodies are oftentimes not applicable to 3D imaging, due to their relatively large size and consequently inadequate tissue penetration and penetration speed. The lack of suitable reagents for 3D histopathology can be overcome by an emerging class of single-domain antibodies, referred to as nanobodies (Nbs), which can facilitate rapid and superior 2D and 3D histological stainings. Here, we report the generation and experimental validation of Nbs directed against the human endothelial cell-selective adhesion molecule (hESAM), which enables spatial visualization of blood vascular networks in whole-mount 3D imaging. After analysis of Nb binding properties and quality, selected Nb clones were validated in 2D and 3D imaging approaches, demonstrating comparable staining qualities to commercially available hESAM antibodies in 2D, as well as rapid and complete staining of entire specimens in 3D. We propose that the presented hESAM-Nbs can serve as novel blood vessel markers in academic research and can potentially improve 3D histopathological diagnostics of entire human tissue specimens, leading to improved treatment and superior patient outcomes. View Full-Text
Keywords: ESAM; nanobodies; single-domain antibodies; light sheet imaging; 3D microscopy; blood vessel marker; histopathology; 3D reconstruction ESAM; nanobodies; single-domain antibodies; light sheet imaging; 3D microscopy; blood vessel marker; histopathology; 3D reconstruction
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MDPI and ACS Style

Hansmeier, N.R.; Büschlen, I.S.; Behncke, R.Y.; Ulferts, S.; Bisoendial, R.; Hägerling, R. 3D Visualization of Human Blood Vascular Networks Using Single-Domain Antibodies Directed against Endothelial Cell-Selective Adhesion Molecule (ESAM). Int. J. Mol. Sci. 2022, 23, 4369. https://doi.org/10.3390/ijms23084369

AMA Style

Hansmeier NR, Büschlen IS, Behncke RY, Ulferts S, Bisoendial R, Hägerling R. 3D Visualization of Human Blood Vascular Networks Using Single-Domain Antibodies Directed against Endothelial Cell-Selective Adhesion Molecule (ESAM). International Journal of Molecular Sciences. 2022; 23(8):4369. https://doi.org/10.3390/ijms23084369

Chicago/Turabian Style

Hansmeier, Nils Rouven, Ina Sophie Büschlen, Rose Yinghan Behncke, Sascha Ulferts, Radjesh Bisoendial, and René Hägerling. 2022. "3D Visualization of Human Blood Vascular Networks Using Single-Domain Antibodies Directed against Endothelial Cell-Selective Adhesion Molecule (ESAM)" International Journal of Molecular Sciences 23, no. 8: 4369. https://doi.org/10.3390/ijms23084369

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