Research on Biomedical Engineering
Research on Biomedical Engineering
Original Article

Generation of 3D ultrasound biomicroscopic images: technique validation and in vivo volumetric imaging of rat lateral gastrocnemius

Martins, Natália Santos da Fonseca; Carneiro, Luisa Tinoco; Dantas, Hugo de Mello; Esperança, Cláudio; Marroquim, Ricardo Guerra; Oliveira, Liliam F.; Machado, João Carlos

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Introduction: Ultrasound biomicroscopy (UBM) is a technique for generating high-resolution images, with frequencies from 20 MHz to 100 MHz. For example, it has been used in animal research related to models of injury and diseases that mimic human conditions. With a three-dimensional ultrasound (3D) image system, an organ can be viewed at various angles and the volume estimated, contributing to an accurate diagnosis. This work refers to the generation of 3D-UBM images, employing a 35 MHz ultrasound system, from multiple two-dimensional (2D) images. Phantoms were used to validate the technique and to determine its reliability of volume measurements. Additionally, the technique was used to obtain 3D images of the rat gastrocnemius muscle. Methods: Four different phantoms were used and ten acquisition sequences of 2D-images acquired for each one. Thereafter, 5 volume segmentations were performed for each acquisition sequence, resulting in 50 measured volumes for each phantom. The physical volumes of all phantoms were used to validate the technique based on the coefficient of variation (CV) and the intraclass correlation coefficient (ICC). Images of the gastrocnemius muscle were acquired and the partial volume quantified. Results: The CV and ICC confirmed the reliability of volume measurements obtained by segmentation. Moreover, cross-sectional 2D images of rat hindlimb were obtained, allowing to identify the gastrocnemius muscle and to partially quantify the muscle volume from 3D images. Conclusion: The results indicated that the technique is valid to generate 3D images and quantify the volume of a muscle compatible with the dimensions of a small animal.


Ultrasound biomicroscopy, 3D image, phantom, gastrocnemius muscle, rat.


Alves KZ, Soletti RC, Britto MA, Matos DG, Soldan M, Borges HL, Machado JC. In vivo endoluminal ultrasound biomicroscopic imaging in a mouse model of colorectal cancer. Academic Radiology 2013; 20(1):90-8. PMid:22959583

Barber L, Barrett R, Lichtwark G. Validation of a freehand 3D ultrasound system for morphological measures of the medial gastrocnemius muscle. Journal of Biomechanics 2009; 42(9):1313-9. PMid:19375081

Chang RF, Wu WJ, Chen DR, Chen WM, Shu W, Lee JH, Jeng LB. 3-D US frame positioning using speckle decorrelation and image registration. Ultrasound in Medicine & Biology 2003; 29(6):801-12. PMid:12837496

Delcker A, Walker F, Caress J, Hunt C, Tegeler C. In vitro measurement of muscle volume with 3-dimensional ultrasound. European Journal of Ultrasound 1999; 9(2):185-90. PMid:10413755

Forsberg F, Berghella V, Merton DA, Rychlak K, Meiers J, Goldberg BB. Comparing image processing techniques for improved 3-dimensional ultrasound imaging. Journal of Ultrasound in Medicine 2010; 29(4):615-9. PMid:20375380.

Foster FS, Hossack J, Adamson SL. Micro-ultrasound for preclinical imaging. Interface Focus. 2011; 1(4):576-601. PMid:22866232

Foster FS, Pavlin CJ, Harasiewicz KA, Christopher DA, Turnbull DH. Advances in ultrasound biomicroscopy. Ultrasound in Medicine & Biology 2000; 26(1):1-27. PMid:10687788

Foster FS, Zhang MY, Zhou YQ, Liu G, Mehi J, Cherin E, Harasiewicz KA, Starkoski BG, Zan L, Knapik DA, Adamson SL. A new ultrasound instrument for in vivo microimaging of mice. Ultrasound in Medicine & Biology 2002; 28(9):1165-72. PMid:12401387

Fry NR, Gough M, McNee AE, Shortland AP. Changes in the volume and length of the medial gastrocnemius after surgical recession in children with spastic diplegic cerebral palsy. Journal of Pediatric Orthopedics 2007; 27(7):769-74. PMid:17878783

Hopkins WG. Measures of reliability in sports medicine and science. Sports Medicine (Auckland, N.Z.) 2000; 30(1):1-15. PMid:10907753

Infantolino BW, Gales DJ, Winter SL, Challis JH. The validity of ultrasound estimation of muscle volumes. Journal of Applied Biomechanics 2007; 23(3):213-7. PMid:18089918.

Kawakami Y, Kanehisa H, Fukunaga T. The relationship between passive ankle plantar flexion joint torque and gastrocnemius muscle and achilles tendon stiffness: implications for flexibility. The Journal of Orthopaedic and Sports Physical Therapy 2008; 38(5):269-76. PMid:18448880

Lieber RL, Fridén J. Functional and clinical significance of skeletal muscle architecture. Muscle & Nerve 2000; 23(11):1647-66.<1647::AID-MUS1>3.0.CO;2-M. PMid:11054744

MacGillivray TJ, Ross E, Simpson HAHRW, Greig CA. 3D freehand ultrasound for in vivo determination of human skeletal muscle volume. Ultrasound in Medicine & Biology 2009; 35(6):928-35. PMid:19185972

Peixinho CC, Martins NSF, de Oliveira LF, Machado JC. Structural adaptations of rat lateral gastrocnemius muscle-tendon complex to a chronic stretching program and their quantification based on ultrasound biomicroscopy and optical microscopic images. Clinical Biomechanics (Bristol, Avon) 2014; 29(1):57-62. PMid:24309012

Peixinho CC, Ribeiro MB, Resende CMC, Werneck-de-Castro JPS, Oliveira LF, Machado JC. Ultrasound biomicroscopy for biomechanical characterization of healthy and injured triceps surae of rats. The Journal of Experimental Biology 2011; 214(Pt 22):3880-6. PMid:22031753

Petter-Puchner A, Gruber-Blum S, Walder N, Fortelny RH, Redl H, Raum K. Ultrasound biomicroscopy (UBM) and scanning acoustic microscopy (SAM) for the assessment of hernia mesh integration: a comparison to standard histology in an experimental model. Hernia 2014; 18(4):579-85. PMid:24346242

Roellig K, Drews B, Goeritz F, Hildebrandt TB. The long gestation of the small naked mole-rat (Heterocephalus glaber Rüppell, 1842) studied with ultrasound biomicroscopy and 3D-ultrasonography. PLoS ONE 2011; 6(3):e17744. PMid:21408185

Ryan LK, Foster FS. Tissue equivalent vessel phantoms for intravascular ultrasound. Ultrasound in Medicine & Biology 1997; 23(2):261-73. PMid:9140183

Shemesh H, Goertz DE, van der Sluis LW, de Jong N, Wu MK, Wesselink PR. High frequency ultrasound imaging of a single-species biofilm. Journal of Dentistry 2007; 35(8):673-8. PMid:17604896

Stachs O, Martin H, Kirchhoff A, Stave J, Terwee T, Guthoff R. Monitoring accommodative ciliary muscle function using three-dimensional ultrasound. Graefes Archive for Clinical and Experimental Ophthalmology 2002; 240(11):906-12. PMid:12486512

truebok _id="b22"truebokTakamasa I, Hiroshi Y, Toshiyuki H. A color atlas of sectional anatomy of the rat. Japan; 2001

alker M 3rd, Campbell BR, Azer K, Tong C, Fang K, Cook JJ, Forrest MJ, Kempadoo K, Wright SD, Saltzman JS, MacIntyre E, Hargreaves R. A novel 3-dimensional micro-ultrasound approach to automated measurement of carotid arterial plaque volume as a biomarker for experimental atherosclerosis. Atherosclerosis 2009; 204(1):55-65. PMid:19135672

Weller R, Pfau T, Ferrari M, Griffith R, Bradford T, Wilson A. The determination of muscle volume with a freehand 3D ultrasonography system. Ultrasound in Medicine & Biology 2007; 33(3):402-7. PMid:17208353
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