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Angiogenesis Analyzer for ImageJ

Saturday 20 October 2012, by Gilles Carpentier

  Table of contents  

This tool was presented at the poster session of the

ImageJ User and Developer Conference 2012


2020 - Angiogenesis Analyzer for ImageJ — A comparative morphometric analysis of “Endothelial Tube Formation Assay” and “Fibrin Bead Assay”.

 Description:


The "Angiogenesis Analyzer" [1] allows analysis of cellular networks. Typically, it can detect and analyze the pseudo vascular organization of endothelial cells cultured in gel medium.

Among the tools used to evaluate the anti-angiogenesis properties of drugs, the most used is the in vitro differentiation of primary endothelial cells culture in gel (Endothelial Tube Formation Assay (ETFA)) [2]. In suitable culture conditions these cells form structures that can branch and mimic a pseudo capillary in vitro formation. At later stage this differentiation can lead to a meshed network from different mesh sizes. Although widely used, the interpretation of this assay still remains a problem, especially to obtain a quantitative evaluation of the vessels-like net organization. The Angiogenesis Analyzer is a simple tool to quantify the ETFA experiment images by extracting characteristic information of the network.

Above: phase contrast image of HUVEC network, analyzed by the Angiogenesis Analyzer for ImageJ. The same analysis can be obtained using a fluorescent staining by calcein.

 Collaborations - image and thematic providers:


- Dr Anaïs Leroy, Dr Carole Brasse-Lagnel and Dr Bruno Gonzalez. Epigenetics and Pathophysiology of Neurodevelopmental Disorders team, INSERM U1245, University of Rouen Normandie, Rouen, France. [3].


- Aude Durant, Dr Olivia Garnier and Dr Isabelle Vilgrain (DR CNRS) CEA Grenoble, France. [4] [5].


- Jerrold R. Turner, Professor of Pathology and Medicine, M.D., Ph.D. and Laura-Sophie Beier, Postdoctoral Research Fellow. Laboratory of Mucosal Barrier Pathology, Department of Pathology, Brigham and Women’s Hospital 77, Avenue Louis Pasteur, Boston, MA. [6] [7].


- Dr Amandine Rovini, Pr Alexis Desmoulière, team, NeurIT Neuropathies périphériques et Innovations Thérapeutiques UR 20218, Faculties of Medicine and Pharmacy, University of Limoges, Limoges, France. [8] [9] [10] [11].


- Fabrice Antigny. Role of ion channels in pulmonary arterial hypertension. INSERM UMR-S999 Pulmonary hypertension: pathophysiology and novel therapies. Hôpital Marie Lannelongue, Le Plessis Robinson, France. [12] [13].


- Elisabeth Vaganay, Sandrine Bretaud, Laurie Nemoze-Billet and Forence Ruggiero, Matrix Biology and Pathology, Institut de Génomique Fonctionnelle de Lyon, ENS de lyon, Université de Lyon, CNRS UMR 5242, France. [14] [15] [16].


- Pr. Richard Cook and Dr. Pedro Bastos, Dental Institute, Guy’s Hospital, King’s College London, UK. [17].


- Dr. Fabien Chevalier, Dr. Ségolène Ferratge and Dr. Georges Uzan. INSERM U1197, Hôpital Paul Brousse, Villejuif. France. [18] [19] [20].


- Dr Cédric ALLIER. CEA-Leti - MINATEC Campus, Grenoble, France. [21].


- Geoffrey Estaban. IPRASENSE, Cap Alpha, Clapiers, France. www.iprasense.com [22].


- Dacha Gholobova, Lieselot Decroix and Prof. dr. ir. Lieven Thorrez. Dept. Development and Regeneration @Kulak, KU Leuven, Kortrijk. Belgium. [23] [24]


- Dr. Arvind P. Pathak and Malvi Hemani, Depts. of Radiology and Oncology, The Johns Hopkins University School of Medicine, Baltimore. USA. Web: www.pathaklab.org. [25]


- Dr Sarah Berndt and Prof Muriel Cuendet. Pharmacognosie, Phytochimie et produits naturels bioactifs. Section des Sciences Pharmaceutiques. Université de Genève, Université de Lausanne Sciences II. Switzerland. [26] [27] [28] [29] [30] [31].


- Dr. Valeria Samarelli and Marco Arese, Associate Professor of Biochemistry, Department of Oncological Sciences - University of Turin - School of Medecine - Institute for Cancer Research and Treatment (IRCC). Italy. [32]

History:

- First version: 15 October 2012

Click the "Update and Version Infos" icon to look for new versions:


Footnotes

[1] Gilles Carpentier. Contribution: Angiogenesis Analyzer. ImageJ News, 5 October 2012.

[2] L. Montesano, R. Orci and P. Vassalli, “In vitro rapid organization of endothelial cells into capillary-like networks is promoted by collagen matrices”, J Cell Biol 97, pp. 1648–52, 1983.

[3] Leroy A, Valentin A, Sautreuil C, Carpentier G, Racine C, Lecointre M, Janin F, Vivien D, Marret S, Picaud S, Gonzalez BJ, Brasse-Lagnel C. Endothelial NMDA Receptor Involvement in Retinal Neurovascular Damage Following Prenatal Alcohol Exposure in a Mouse Model. J Neurosci. 2026 Apr 22;46(16):e1419252026. doi:10.1523/JNEUROSCI.1419-25.2026. Abstract

[4] Aude Durand, Amandine Pitaval, Pierre Bleuet, Gilles Carpentier, Sarah Berndt, Quentin Perrier, Emily Tubbs, Sandrine Lablanche, Marie-Line Cosnier, Cécile Moro. Evaluation of photobiomodulation effects on vascular network using physiological model. MPS World Summit 2023 - June 26 – June 30, 2023 Berlin, Germany.

[5] Garnier O, Jeanneret F, Durand A, Fertin A, Berndt S, Carpentier G, Battail C, Martin DK, Vilgrain I. Efficient Fine-Tuning of Endothelial Gene Expression by a Single Tyrosine (Y685) to Phenylalanine Point Mutation in the VE-Cadherin Gene. Arterioscler Thromb Vasc Biol. 2025 Aug 14. doi: 10.1161/ATVBAHA.125.323129. Online ahead of print. PMID: 40808655. Abstract.

[6] Laura-Sophie Beier, Xian Shi, Gilles Carpentier, Patrick M. Marsch, Michael Grey, Shikha Nangia, Jerrold R. Turner. Molecular regulation of tight junction strand assembly and channel function. American Physiology Summit 2025 APRIL 24 – 27, 2025, BALTIMORE, USA.

[7] L. Beier, P. M. Marsch, G. Carpentier, S. Nangia, J. R. Turner. Integrating structural determinants of claudin-2 with strand formation and channel function. Cell Bio 23 ASCB-EMBO meeting 2023 - December 2-6, 2023 Boston, Massachusetts USA. Abstract.

[8] Sbeih M., Serbier M., Carpentier G., Duciel L., Des Courtils C., Laloze J., Desmoulière A., Rovini A. Comparative Evaluation Of Calcium Alginate And Cmc-Based Dressings On Sensory Neuron Function And Regeneration. NeuroFrance 2025. 14–16 mai 2025, Montpellier, France.

[9] Rovini, A; Casas, E; Siadous, R; Guerber, L; Bellenguez, M; Carpentier, G; Leng, J; Bordes, S; Closs, B; Berthod, F; Amédée, J; Desmoulière, A. Schwann cells contribution to neurovascular interactions in modelling cutaneous innervation. Annual Meeting of the Peripheral-Nerve-Society (PNS). Journal of The Peripheral Nervous System Supplement Meeting AbstractP 353. Volume 29 150-S151. Montreal, Canada, Jun 22-25, 2024.

[10] Laloze J, Lacoste M, Marouf F, Carpentier G, Vignaud L, Chaput B, Varin A, Desmoulière A, Rovini A. Specific Features of Stromal Cells Isolated from the Two Layers of Subcutaneous Adipose Tissue: Roles of Their Secretion on Angiogenesis and Neurogenesis. J Clin Med. 2023 Jun 22;12(13):4214. doi: 10.3390/jcm12134214. Abstract.

[11] Serbier M, Carpentier G, Sbeih M, Duciel L, Courtils CD, Desmoulière A, Rovini A. Differential Effects of Calcium Alginate and Carboxymethylcellulose Wound Dressing Extracts on Human Sensory Neuron Regeneration and Secretome. Wound Repair Regen. 2026 May-Jun;34(3):e70182. doi: 10.1111 wrr.70182. Abstract.

[12] Le Ribeuz H, Saint-Martin Willer A, Chevalier B, Sancho M, Masson B, Eyries M, Jung V, Guerrera IC, Dutheil M, El Jekmek K, Laubry L, Carpentier G, Perez-Vizcaino F, Tu L, Guignabert C, Chaumais MC, Péchoux C, Humbert M, Hinzpeter A, Mercier O, Capuano V, Montani D, Antigny F. Role of KCNK3 Dysfunction in Dasatinib-associated PAH and Endothelial Cell Dysfunction. Am J Respir Cell Mol Biol. 2024 Mar 28. doi: 10.1165/rcmb.2023-0185OC. Epub ahead of print. PMID: 38546978. Abstract.

[13] Saint-Martin Willer A, Masson B, Laubry L, Dutheil M, El Jekmek K, Carpentier G, Gourmelon M, Sabourin J, Humbert M, Mercier O, Ruffenach G, Montani D, Capuano V, Antigny F. TRPC channels contribute to endothelial dysfunction in pulmonary arterial hypertension. Respir Res. 2025 Dec 12;26(1):345. doi: 10.1186/s12931-025-03376-6. Abstract.

[14] Nemoz-Billet L, Balland M, Gilquin L, Gillet B, Stévant I, Guillon E, Hughes S, Carpentier G, Vaganay E, Sohm F, Misiak V, Gonzalez-Melo MJ, Koch M, Ghavi-Helm Y, Bretaud S, Ruggiero F. Dual topologies of myotomal collagen XV and Tenascin C act in concert to guide and shape developing motor axons. Proc Natl Acad Sci U S A. 2024 Mar 26;121(13):e2314588121. doi: 10.1073/pnas.2314588121. Epub 2024 Mar 19. PMID: 38502691; PMCID: PMC10990108. Abstract.

[15] Nemoz-Billet L., E. Vaganay E., Guillon E., Gillet B., Balland M., Carpentier G., Stévant I., Ruggiero F., Bretaud S. The myotomal collagen XV-B is a key player in motor axon development in zebrafish. 19èmes Journées de la Société Française de Myologie. Nov 23-25, Toulouse, 2022.

[16] Jia T, Vaganay E, Carpentier G, Coudert P, Guzman-Gonzales V, Manuel R, Eymin B, Coll JL, Ruggiero F. A collagen Vα1-derived fragment inhibits FGF-2 induced-angiogenesis by modulating endothelial cells plasticity through its heparin-binding site. Matrix Biol. 2020 Dec;94:18-30. doi: 10.1016/j.matbio.2020.07.001. Abstract.

[17] Bastos P, Carpentier G, Patel V, Papy-Garcia D, Watson T, Cook R. Real-Time Optical Vascular Imaging, a new method for the diagnosis and monitoring of oral diseases.
. J Microsc. 2020 Oct 29. doi: 10.1111/jmi.12975. Online ahead of print. Abstract.

[18] - Carpentier G, Berndt S, Ferratge S, Rasband W, Cuendet M, Uzan G, Albanese P. Angiogenesis Analyzer for ImageJ - A comparative morphometric analysis of "Endothelial Tube Formation Assay" and "Fibrin Bead Assay". Sci Rep. 2020 Jul 14;10(1):11568. doi: 10.1038/s41598-020-67289-8. Abstract.

[19] Ferratge S, Ha G, Carpentier G, Arouche N, Bascetin R, Muller L, Germain S, Uzan G. Initial clonogenic potential of human endothelial progenitor cells is predictive of their further properties and establishes a functional hierarchy related to immaturity. Stem Cell Res. 2017 May;21:148-159. Abstract.

[20] Chevalier F, Lavergne M, Negroni E, Ferratge S, Carpentier G, Gilbert-Sirieix M, Siñeriz F, Uzan G, Albanese P. Glycosaminoglycan mimetic improves enrichment and cell functions of human endothelial progenitor cell colonies. Stem Cell Res. May;12(3):703-15, 2014. Abstract.

[21] Allier C, Esteban G, Carpentier G, Van Der Sanden B, Vinjimore Kesavan S, Coutard J-G, Bordy T, Momey F, Herve L, Hamard L, Di Natale C, Wion D, Dinten J-M. Video Lensfree Microscopy for the Quantification of Endothelial Cellular Network Formation. 6ème Congrès de la Société Française d’Angiogenèse. Paris Collège de France – Septembre 24-25, 2015.

[22] Allier C, Esteban G, Carpentier G, Van Der Sanden B, Vinjimore Kesavan S, Coutard J-G, Bordy T, Momey F, Herve L, Hamard L, Di Natale C, Wion D, Dinten J-M. Video Lensfree Microscopy for the Quantification of Endothelial Cellular Network Formation. 6ème Congrès de la Société Française d’Angiogenèse. Paris Collège de France – Septembre 24-25, 2015.

[23] - Gholobova D, Terrie L, Mackova K, Desender L, Carpentier G, Gerard M, Hympanova L, Deprest J, Thorrez L. Functional evaluation of prevascularization in one-stage versus two-stage tissue engineering approach of human bio-artificial muscle. Biofabrication. 2020 Jun 12;12(3):035021. doi: 10.1088/1758-5090/ab8f36. PMID: 32357347. Abstract.

[24] Gholobova D, Decroix L, Van Muylder V, Desender L, Gerard M, Carpentier G, Vandenburgh H, Thorrez L. Endothelial Network Formation Within Human Tissue-Engineered Skeletal Muscle. Tissue Eng Part A. 2015 Sep 1. Abstract.

[25] Fortenberry YM, Brandal SM, Carpentier G, Hemani M, Pathak AP. Intracellular Expression of PAI-1 Specific Aptamers Alters Breast Cancer Cell Migration, Invasion and Angiogenesis. PLoS One. 2016 Oct 18;11(10):e0164288. Abstract.

[26] - Carpentier G, Berndt S, Ferratge S, Rasband W, Cuendet M, Uzan G, Albanese P. Angiogenesis Analyzer for ImageJ - A comparative morphometric analysis of "Endothelial Tube Formation Assay" and "Fibrin Bead Assay". Sci Rep. 2020 Jul 14;10(1):11568. doi: 10.1038/s41598-020-67289-8. Abstract.

[27] Berndt S, Carpentier G, Turzi A, Borlat F, Cuendet M, Modarressi A. Angiogenesis Is Differentially Modulated by Platelet-Derived Products. Biomedicines. 2021 Mar 4;9(3):251. doi: 10.3390/biomedicines9030251. Abstract.

[28] Sakr OS, Berndt S, Carpentier G, Cuendet M, Jordan O, Borchard G. Arming embolic beads with anti-VEGF antibodies and controlling their release using LbL technology. J Control Release. 2016 Feb 28;224:199-207. Abstract.

[29] Queiroz MMF, Monteillier A, Berndt S, Marcourt L, Franco ES, Carpentier G, Nejad Ebrahimi S, Cuendet M, Bolzani VDS, Maia MBS, Queiroz EF, Wolfender JL. NF-κB and Angiogenesis Inhibitors from the Aerial Parts of Chresta martii. J Nat Prod. 2018 Aug 24;81(8):1769-1776. Abstract.

[30] Issa ME, Berndt S, Carpentier G, Pezzuto JM, Cuendet M. Bruceantin inhibits multiple myeloma cancer stem cell proliferation. Cancer Biol There. 2016 Jul 19:0. Abstract.

[31] Berndt S, Issa ME, Carpentier G, Cuendet M. A Bivalent Role of Genistein in Sprouting Angiogenesis. Planta Med. 2018 Mar 14. doi: 10.1055/a-0587-5991. Abstract.

[32] Samarelli AV, Riccitelli E, Bizzozero L, Nunes Silveira T, Seano G, Pergolizzi M, Vitagliano G, Cascone I, Carpentier G, Bottos A, Primo L, Bussolino F, Arese M. Neuroligin 1 induces blood vessels maturation by cooperating with the α6 integrin. J Biol Chem. May 23. pii: jbc.M113.530972. 2014. Abstract

logoij ImageJ (http://rsb.info.nih.gov/ij/) is a public domain Java image processing program inspired by NIH Image for the Macintosh. It runs, either as an online applet or as a downloadable application, on any computer with a Java 1.1 or later virtual machine. Downloadable distributions are available for Windows, Mac OS, Mac OS X and Linux. The author, Wayne Rasband (wayne@codon.nih.gov), is at the Research Services Branch, National Institute of Mental Health, Bethesda, Maryland, USA.
Gilles Carpentier, Faculte des Sciences et Technologie,
Universite Paris Est Creteil Val-de-Marne, France.

Special thanks to Alessandra Albano for the English correction of these sites.
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