We thank the nurses (appearing on film) for supporting the neonates. 
We thank J. Hagoort, MA, linguist, Department of Pediatric Surgery, Erasmus MC-Sophia Children&#8217;s Hospital, Rotterdam, the Netherlands, for reading and correcting the manuscript.

NOTA: Este protocolo se ajusta a las directrices del comité de ética de investigación en humanos local. 1. Consideraciones generales NOTA: Consideraciones generales sobre los equipos, almacenamiento y seguridad de los datos se tratan en la Discusión. <ol><li> Obtener imágenes con una alta resolución, en tiempo real, móvil máquina de ultrasonido en 2D con múltiples transductores con una banda de frecuencias (ver Discusión). Típicamente, obtener imágenes de buena calidad usando una sonda con una frecuencia de 7.5 a 8.5 MHz. </li></ol> 2. Preparación del Examen CUS <ol><li> Programar el examen CUS de modo que no coincide con otros procedimientos tales como el muestreo de sangre. </li><li> Asegúrese de que un trabajador de la salud o un padre está disponible para apoyar y / o confort del neonato durante el examen, utilizando estrategias como las que de acuerdo con los principios del Programa de Evaluación del recién nacido 6 Individualizado de Atención y Desarrollo &lt;/ Sup&gt;. </li></ol> 3. Examen través fontanela anterior <ol><li> Instale la máquina de ultrasonido a lo largo de la incubadora o cuna. </li><li> Aplicar gel transductor a la sonda para asegurar un buen contacto entre la sonda y la piel. Considere el calentamiento del gel antes de su uso. </li><li> Comience imágenes a través de la fontanela anterior con una sonda convexa en modo B. Coloque la sonda en el medio de la fontanela con el marcador en la sonda se volvió hacia el lado derecho de la neonato. El lado izquierdo del cerebro se mostrará a continuación en el lado derecho del monitor. NOTA: Imaging a través de la fontanela anterior se puede realizar con el recién nacido en cualquier posición 3. Para fines de investigación, puede ser necesario para luchar por una posición de cabeza de serie. <ol><li> Graba imágenes en al menos cinco coronal y cinco planos sagital. En la primera imagen ajuste de profundidad, ajustes de compensación de ganancia de ganancia y tiempo para producir una imagen que llena el sector, que contiene los contornos craneales, evitaring imágenes demasiado claras u oscuras y con el objetivo de un equilibrio entre los reflejos de las estructuras cercanas y profundas. </li><li> Planos coronal NOTA: Trate de obtener imágenes perfectamente simétricas. Cuando se sospecha lesiones cerca de la convexidad del lóbulo frontal, considere la grabación secciones coronales oblicuas específicas, de modo que un hemisferio se muestra en mayor detalle (Figura 1). <ol><li> Para la imagen prefrontal coronal, ángulo hacia delante de la sonda Para visualizar los lóbulos frontales, anterior a los cuernos frontales de los ventrículos laterales a nivel de los surcos olfativo. </li><li> Para la imagen coronal a nivel de Monro, ángulo de la sonda para visualizar la sección anterior coronal a la tela coroidea para representar los cuernos frontales de los ventrículos laterales, cavum septum pellucidum, cuerpo calloso, cinguli surco. Nota la ecogenicidad de partes de los ganglios basales. </li><li> Para la imagen coronal al nivel de tálamo, el ángulo de la sonda hacia atrás para identificar la tardefisuras ral, tela coroidea en el techo del tercer ventrículo y lóbulos temporales. Tenga en cuenta la ecogenicidad del tálamo (especialmente los núcleos ventrolateral) en relación con los ganglios basales. Tenga en cuenta que la lesión de la red en pulvinar puede ser visualizado en una sección coronal adicional justo en frente de las aurículas. </li><li> Para la imagen coronal a nivel de las aurículas, los ventrículos laterales visualizar en el nivel del plexo coroideo. Identificar los lóbulos temporales y hemisferios del cerebelo. Tenga en cuenta la ecogenicidad de la sustancia blanca periventricular en comparación con el plexo coroideo. Comparar la radiación óptica con las áreas hiperecoicas normales anteriores y laterales a las aurículas en los recién nacidos prematuros. </li><li> Para la imagen parieto-occipital coronal, ángulo de la sonda hacia atrás al nivel del surco parieto-occipital para identificar los lóbulos parietal y occipital. </li></ol></li><li> Sagital <ol><li> Girar la sonda de 90 ° con el marcador en la sonda orientada hacia la cara del neonato. The parte anterior del cerebro se muestra en el lado izquierdo del monitor. Graba imágenes en el plano de las siguientes estructuras (Figura 2). </li><li> Para la imagen sagital medio, visualizar el cuerpo calloso, cavum septum pellucidum (CSP), tercero y cuarto ventrículo, vermis, cisterna magna, protuberancia y el mesencéfalo. Cabe destacar la presencia de cavum Vergae y velo cavum interpositi 7. </li><li> Para la imagen parasagital a través de uno ovoide gangliothalamic (por ejemplo, el derecho), el ángulo de la sonda de lado para una vista parasagital a través del ventrículo lateral. Identificar el plexo coroideo y anote la ecogenicidad del tálamo y los ganglios basales. El lado escaneada para aviones parasagitales debe indicarse adecuadamente con herramientas de texto. </li><li> Para el parasagital imagen insular, el ángulo de la sonda más lateral a través de la ínsula. Identificar la fisura lateral y frontal-, temporalidad, parietal- y lóbulos occipitales. </li><li> Imágenes parasagitales Repita para tél lado contralateral (es decir, la izquierda). </li></ol></li><li> Doppler Color <ol><li> Continuar imágenes a través de la fontanela anterior con una sonda convexa utilizando Doppler color. Considere la posibilidad de evaluar las velocidades de flujo en las arterias cerebrales y las venas y la obtención de los índices derivados. NOTA: índice de resistencia (RI) se define como la velocidad pico sistólica - velocidad diastólica final / velocidad pico sistólica. RI es independiente del ángulo, los valores de velocidad absolutos no son 8-10. RI no es similar en las arterias de diferente calibre. Las mediciones en serie sólo son útiles si se realiza en la misma ubicación exacta del mismo recipiente. </li></ol></li><li> Graba imágenes en planos coronal de los siguientes buques (Figura 3): <ol><li> Visualizar los senos transversales a nivel del cerebelo. Si sólo uno o ninguno los senos transversal se visualiza, intente reducir la frecuencia de repetición de impulsos (PRF). Si entonces todavía solamente uno o ninguno de los senos transversalespueden ser identificados a través de la fontanela anterior, utilice una sonda lineal de alta frecuencia para la visualización a través de la fontanela mastoidea (ver sección 4.4.2). </li><li> Visualice el polígono de Willis con arterias carótidas internas, las arterias cerebrales medias y anterior arterias cerebrales a nivel de las astas frontales de los ventrículos laterales. Distinguir las arterias cerebrales anteriores izquierdo y derecho es a menudo difícil, pero no suele ser necesario. Identificar los candelabros del estriado de las arterias. </li><li> Ángulo de la sonda hacia atrás para visualizar la arteria basilar con venas yugulares adyacentes. </li><li> Ángulo aún más hacia atrás para visualizar las venas cerebrales y talamostriada internos. </li></ol></li><li> Grabar una imagen en el plano sagital de una arteria cerebral anterior (Figura 4). Evaluar la velocidad del flujo y RI en una parte específica de este buque (por lo general por debajo de la rodilla del cuerpo calloso). Cerca de las velocidades de la línea media en la vena cerebral interna pueden sermedido fácilmente. </li><li> El uso de una sonda lineal de alta frecuencia en un plano coronal en la fontanela anterior, identificar el seno sagital superior. Si esto falla, reducir la cantidad de presión aplicada con la sonda a la fontanela. NOTA: La sonda lineal se puede utilizar para la visualización detallada de las estructuras superficiales (meninges, aracnoideo y el espacio subdural, la corteza). Vasos son tangenciales en el espacio subaracnoideo. Idealmente, se realizará de formación de imágenes Doppler, como se describe en los pasos anteriores durante el primer examen CUS del neonato. Durante el examen de seguimiento algunos de los pasos que se pueden saltar. En caso de Doppler trombosis seno-venosa cerebral se sospecha como se describe en los pasos 3.3.5.1, 3.3.7 y 4.4.2 se debe realizar. </li></ol></li></ol> 4. Examen través de Acústica de Windows Alternativa <ol><li> A continuación, continúe el examen a través de ventanas acústicas alternativas. </li><li> Considere la posibilidad de grabar imágenes a través de la lambdoide (posterior) Fontanela utilizando una sonda convexa (Figura 5). La fontanela posterior se encuentra en la unión de las suturas sagital y lambdoid 3,11. Imagen a través de la fontanela posterior colocando al recién nacido en posición de decúbito lateral. NOTA: En muchos bebés prematuros imágenes satisfactorias también se pueden obtener a través de la cara posterior de la sutura sagital con el niño en una posición supina 3. <ol><li> Coloque la sonda en el medio de la fontanela posterior para una vista sagital. Ángulo de la sonda ligeramente fuera de la línea media para identificar el cuerpo del ventrículo lateral y su cuerno occipital. Gire la sonda de aproximadamente 90 ° para obtener una vista coronal. Identificar los cuernos occipitales de los ventrículos laterales. </li></ol></li><li> Considere la posibilidad de grabar imágenes a través de la (temporal) ventana lateral utilizando una sonda convexa o lineal por encima de la oreja (Figura 6). <ol><li> Si es necesario, obtener imágenes a través de la ventana lateral para permitir unavista detallada del tronco cerebral 12. Coloque la sonda horizontalmente por encima y ligeramente por delante de la oreja. Mueva la sonda hasta que se visualizan los pedúnculos cerebrales. NOTA: Otras estructuras que pueden ser identificados son el tercer ventrículo, el acueducto y los lóbulos temporales. El uso de Doppler color, el círculo de Willis se puede visualizar. </li></ol></li><li> Graba imágenes a través de la fontanela mastoidea (Figura 7). La fontanela mastoidea se encuentra detrás de la oreja, en el cruce de la temporal, parietal y occipital 4. Imagen a través de la fontanela mastoidea colocando al recién nacido en una posición de decúbito lateral 3. NOTA: En nuestra experiencia, los recién nacidos a menudo muestran signos de malestar cuando se obtienen imágenes a través de la fontanela mastoidea. Por lo tanto, sería mejor hacer esto después de la formación de imágenes a través de la fontanela anterior y otras ventanas acústicas. Se postula que este malestar podría ser causado por el mecanismo de la respuesta auditiva a pulses de energía de radiofrecuencia 13. <ol><li> Imagen a través de la fontanela mastoidea utilizando una sonda convexa. Coloque la sonda en paralelo a la oreja para obtener una vista coronal. Barrer la sonda de ida y vuelta para identificar los cerebelo hemisferios, vermis, tercero y cuarto ventrículo, protuberancia y cisterna magna. En pequeños prematuros hemisferio cerebeloso contralateral puede estar bien representado. </li><li> Imagen a través de la fontanela mastoidea utilizando una sonda lineal. Si (uno de) los senos transversales no pueden ser identificados a través de la fontanela anterior, utilice una sonda lineal de alta frecuencia para la visualización a través de la fontanela mastoidea. Coloque la sonda paralela al lóbulo de la oreja para obtener una vista coronal. <ol><li> Identificar el hemisferio cerebeloso y cuarto ventrículo. Utilizando Doppler color, identificar el transverso y sigmoideo seno, seno tentorial y venas emisarias. </li></ol></li></ol></li><li> Considere la posibilidad de visualización adicional de la fosa posterior través foramen magnum 14. </li></ol>

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The authors have nothing to disclose.

Describimos y demostrar un enfoque estado-of-the-art para CUS Doppler neonatal. En manos expertas, esta es una excelente herramienta para, junto a la cama de serie de imágenes del cerebro neonatal seguro. En muchas UCIN no se utilizan de forma óptima las posibilidades descritas. Adición de estudios Doppler permite la detección de la permeabilidad de las arterias intracraneales y las venas. Las velocidades de flujo pueden ser evaluados y los índices obtenidos. Doppler CUS permite la detección de trombosis seno-veno...

Since its clinical introduction in the late 1970&#8217;s cranial ultrasound (CUS) has been widely used for detecting congenital anomalies and acquired brain lesions during the neonatal period. In many neonatal intensive care units (NICUs), CUS has become indispensable in the care for critically ill neonates. Major advantages are its relatively low cost and the fact that it can be performed at bedside, even when a patient is unstable. In addition it is radiation-free and allows for serial imaging. Another technique often used for neuroimaging in critically ill neonates is magnetic resonance imaging (MRI). MRI provides excellent image quality, but its clinical use in NICU&#8217;s is currently limited because of logistic and safety issues1. 
Over time, quality of CUS has drastically improved, with advancing technique leading to higher resolution, faster image processing and digital display and back-up. Important brain structures can be adequately visualized using optimal settings. Traditionally, images are obtained through the anterior fontanel. This approach is less suitable for evaluation of infratentorial structures because they are located far away from the transducer and the highly echoic tentorium impedes their assessment. Use of high-frequency linear transducers through alternative acoustic windows and adapted settings also provides access to these brain regions. Examples of these supplemental acoustic windows are the lambdoid (posterior), mastoid and lateral (temporal) fontanels. So far, however, only few NICUs use these additional acoustic windows routinely2-5. Doppler techniques can be used for screening patency of intracranial vessels. Flow velocities and indices in cerebral arteries can also be obtained. Some manufacturers now provide hardware to visualize flow around 2 cm/sec (Raets, et al., unpublished data). Small vessels are well displayed: medullary trunks and channels, subependymal veins tributing to the thalamostriate veins, and perforator arteries. 
We present our approach of neonatal CUS, focusing on the use of different transducers, multiple acoustic windows and Doppler techniques. Neonatologists and radiologists use this approach in daily clinical practice but is also suitable for research purposes. In the practical part of the video we demonstrate bedside use in the NICU.

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			<td height="42" style="height:42px;width:216px;">Name of Material/ Equipment</td>
			<td style="width:191px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:167px;">Comments/Description</td>
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			<td height="21" style="height:21px;width:216px;">MyLab 70</td>
			<td style="width:191px;">Esaote (Genoa, Italy)</td>
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			<td>Ultrasound system</td>
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state of the art cranial ultrasound imaging in neonates

Cranial ultrasound (CUS) is a reputable tool for brain imaging in critically ill neonates. It is safe, relatively cheap and easy to use, even when a patient is unstable. In addition it is radiation-free and allows serial imaging. CUS possibilities have steadily expanded. However, in many neonatal intensive care units, these possibilities are not optimally used. We present a comprehensive approach for neonatal CUS, focusing on optimal settings, different probes, multiple acoustic windows and Doppler techniques. This approach is suited for both routine clinical practice and research purposes. In a live demonstration, we show how this technique is performed in the neonatal intensive care unit. Using optimal settings and probes allows for better imaging quality and improves the diagnostic value of CUS in experienced hands. Traditionally, images are obtained through the anterior fontanel. Use of supplemental acoustic windows (lambdoid, mastoid, and lateral fontanels) improves detection of brain injury. Adding Doppler studies allows screening of patency of large intracranial arteries and veins. Flow velocities and indices can be obtained. Doppler CUS offers the possibility of detecting cerebral sinovenous thrombosis at an early stage, creating a window for therapeutic intervention prior to thrombosis-induced tissue damage. Equipment, data storage and safety aspects are also addressed.

Ejemplos de formación de imágenes hecha de acuerdo con el protocolo descrito se presentan en las Figuras 1 - 7. Las imágenes deben ser cuidadosamente interpretados por un observador experimentado. Imagen simétrica es necesaria para una adecuada interpretación de las imágenes coronales realizadas a través de la fontanela anterior (Figura 1). Cualquier lesión sospechosa debe ser visualizado en tanto a como (medio) plano sagital coronal o por la visualización a tr...

Watch this Scientific Journal Video about State of the Art Cranial Ultrasound Imaging in Neonates at JoVE.com

State of the Art Cranial Ultrasound Imaging in Neonates

Cranial ultrasound (CUS) is a valuable tool for brain imaging in critically ill neonates. This video shows a comprehensive approach for neonatal (Doppler) CUS for both clinical and research purposes, including a bedside demonstration of the technique.

Estado del Arte craneal imágenes por ultrasonido en neonatos

Cranial ultrasound (CUS) is a reputable tool for brain imaging in critically ill neonates. It is safe, relatively cheap and easy to use, even when a ...

state-of-the-art-cranial-ultrasound-imaging-in-neonates

Research

JoVE Journal

Medicine

23.8K vistas.  Erasmus MC-Sophia Children's Hospital. Cranial ultrasound (CUS) is a valuable tool for brain imaging in critically ill neonates. This video shows a comprehensive approach for neonatal (Doppler) CUS for both clinical and research purposes, including a bedside demonstration of the technique. 

Video: State of the Art Cranial Ultrasound Imaging in Neonates

Dual-mode Imaging of Cutaneous Tissue Oxygenation and Vascular Function

Accurate assessment of cutaneous tissue oxygenation and vascular function is important for appropriate detection, staging, and treatment of many health disorders such as chronic wounds. We report the development of a dual-mode imaging system for non-invasive and non-contact imaging of cutaneous tissue oxygenation and vascular function. The imaging system integrated an infrared camera, a CCD camera, a liquid crystal tunable filter and a high intensity fiber light source. A Labview interface was programmed for equipment control, synchronization, image acquisition, processing, and visualization. Multispectral images captured by the CCD camera were used to reconstruct the tissue oxygenation map. Dynamic thermographic images captured by the infrared camera were used to reconstruct the vascular function map. Cutaneous tissue oxygenation and vascular function images were co-registered through fiduciary markers. The performance characteristics of the dual-mode image system were tested in humans.

A dual-mode imaging system was developed for non-contact assessment of cutaneous tissue oxygenation and vascular function.

De modo dual de imágenes de la oxigenación de los tejidos cutáneos y la función vascular

Accurate assessment of cutaneous tissue oxygenation and vascular function is important for appropriate detection, staging, and treatment of many health ...

Investigación

Medicina

Murine Echocardiography and Ultrasound Imaging

Rodent models of cardiac pathophysiology represent a valuable research tool to investigate mechanism of disease as well as test new therapeutics.1 Echocardiography provides a powerful, non-invasive tool to serially assess cardiac morphometry and function in a living animal.2 However, using this technique on mice poses unique challenges owing to the small size and rapid heart rate of these animals.3 Until recently, few ultrasound systems were capable of performing quality echocardiography on mice, and those generally lacked the image resolution and frame rate necessary to obtain truly quantitative measurements. Newly released systems such as the VisualSonics Vevo2100 provide new tools for researchers to carefully and non-invasively investigate cardiac function in mice. This system generates high resolution images and provides analysis capabilities similar to those used with human patients. Although color Doppler has been available for over 30 years in humans, this valuable technology has only recently been possible in rodent ultrasound.4,5 Color Doppler has broad applications for echocardiography, including the ability to quickly assess flow directionality in vessels and through valves, and to rapidly identify valve regurgitation. Strain analysis is a critical advance that is utilized to quantitatively measure regional myocardial function.6 This technique has the potential to detect changes in pathology, or resolution of pathology, earlier than conventional techniques. Coupled with the addition of three-dimensional image reconstruction, volumetric assessment of whole-organs is possible, including visualization and assessment of cardiac and vascular structures. Murine-compatible contrast imaging can also allow for volumetric measurements and tissue perfusion assessment.

This video demonstrates use of a rail-mounted high-frequency ultrasound probe to perform echocardiography on an anesthetized mouse. The methods describe both conventional two-dimensional and M-mode measurements of cardiac function in addition to newer, more powerful tools such as color Doppler, strain analysis, as well as general and targeted contrast imaging.

Ecocardiografía murino y la ecografía

Rodent models of cardiac pathophysiology represent a valuable research tool to investigate mechanism of disease as well as test new therapeutics.1  ...

MRI-guided Disruption of the Blood-brain Barrier using Transcranial Focused Ultrasound in a Rat Model

Focused ultrasound (FUS) disruption of the blood-brain barrier (BBB) is an increasingly investigated technique for circumventing the BBB1-5. The BBB is a significant obstacle to pharmaceutical treatments of brain disorders as it limits the passage of molecules from the vasculature into the brain tissue to molecules less than approximately 500 Da in size6. FUS induced BBB disruption (BBBD) is temporary and reversible4 and has an advantage over chemical means of inducing BBBD by being highly localized. FUS induced BBBD provides a means for investigating the effects of a wide range of therapeutic agents on the brain, which would not otherwise be deliverable to the tissue in sufficient concentration. While a wide range of ultrasound parameters have proven successful at disrupting the BBB2,5,7, there are several critical steps in the experimental procedure to ensure successful disruption with accurate targeting. This protocol outlines how to achieve MRI-guided FUS induced BBBD in a rat model, with a focus on the critical animal preparation and microbubble handling steps of the experiment.

Microbubble-mediated focused ultrasound disruption of the blood-brain barrier (BBB) is a promising technique for non-invasive targeted drug delivery in the brain1-3. This protocol outlines the experimental procedure for MRI-guided transcranial BBB disruption in a rat model.

Resonancia magnética guiada por disrupción de la barrera sangre-cerebro con ultrasonido transcraneal Enfocado en un modelo de rata

Focused ultrasound (FUS) disruption of the blood-brain barrier (BBB) is an increasingly investigated technique for circumventing the BBB1-5. The BBB is a ...

Multi-modal Imaging of Angiogenesis in a Nude Rat Model of Breast Cancer Bone Metastasis Using Magnetic Resonance Imaging, Volumetric Computed Tomography and Ultrasound

Angiogenesis is an essential feature of cancer growth and metastasis formation. In bone metastasis, angiogenic factors are pivotal for tumor cell proliferation in the bone marrow cavity as well as for interaction of tumor and bone cells resulting in local bone destruction. Our aim was to develop a model of experimental bone metastasis that allows in vivo assessment of angiogenesis in skeletal lesions using non-invasive imaging techniques.
For this purpose, we injected 105 MDA-MB-231 human breast cancer cells into the superficial epigastric artery, which precludes the growth of metastases in body areas other than the respective hind leg1. Following 25-30 days after tumor cell inoculation, site-specific bone metastases develop, restricted to the distal femur, proximal tibia and proximal fibula1. Morphological and functional aspects of angiogenesis can be investigated longitudinally in bone metastases using magnetic resonance imaging (MRI), volumetric computed tomography (VCT) and ultrasound (US).
MRI displays morphologic information on the soft tissue part of bone metastases that is initially confined to the bone marrow cavity and subsequently exceeds cortical bone while progressing. Using dynamic contrast-enhanced MRI (DCE-MRI) functional data including regional blood volume, perfusion and vessel permeability can be obtained and quantified2-4. Bone destruction is captured in high resolution using morphological VCT imaging. Complementary to MRI findings, osteolytic lesions can be located adjacent to sites of intramedullary tumor growth. After contrast agent application, VCT angiography reveals the macrovessel architecture in bone metastases in high resolution, and DCE-VCT enables insight in the microcirculation of these lesions5,6. US is applicable to assess morphological and functional features from skeletal lesions due to local osteolysis of cortical bone. Using B-mode and Doppler techniques, structure and perfusion of the soft tissue metastases can be evaluated, respectively. DCE-US allows for real-time imaging of vascularization in bone metastases after injection of microbubbles7.
In conclusion, in a model of site-specific breast cancer bone metastases multi-modal imaging techniques including MRI, VCT and US offer complementary information on morphology and functional parameters of angiogenesis in these skeletal lesions.

In the pathogenesis of bone metastasis, angiogenesis is a crucial process and therefore represents a target for imaging and therapy. Here, we present a rat model of site-specific breast cancer bone metastasis and describe strategies to non-invasively image angiogenesis in vivo using magnetic resonance imaging, volumetric computed tomography and ultrasound.

Multimodal de imágenes de la angiogénesis en un modelo de rata desnuda de las metástasis óseas del cáncer de pecho utilizando imágenes de resonancia magnética, tomografía computarizada y la ecografía volumétrica

Angiogenesis is an essential feature of cancer growth and metastasis formation. In bone metastasis, angiogenic factors are pivotal for tumor cell ...

Imaging Glioma Initiation In Vivo Through a Polished and Reinforced Thin-skull Cranial Window

Glioma is the one of the most lethal forms of human cancer. The most effective glioma therapy to date-surgery followed by radiation treatment-offers patients only modest benefits, as most patients do not survive more than five years following diagnosis due to glioma relapse 1,2. The discovery of cancer stem cells in human brain tumors holds promise for having an enormous impact on the development of novel therapeutic strategies for glioma 3. Cancer stem cells are defined by their ability both to self-renew and to differentiate, and are thought to be the only cells in a tumor that have the capacity to initiate new tumors 4. Glioma relapse following radiation therapy is thought to arise from resistance of glioma stem cells (GSCs) to therapy 5-10. In vivo, GSCs are shown to reside in a perivascular niche that is important for maintaining their stem cell-like characteristics 11-14. Central to the organization of the GSC niche are vascular endothelial cells 12. Existing evidence suggests that GSCs and their interaction with the vascular endothelial cells are important for tumor development, and identify GSCs and their interaction with endothelial cells as important therapeutic targets for glioma. The presence of GSCs is determined experimentally by their capability to initiate new tumors upon orthotopic transplantation 15. This is typically achieved by injecting a specific number of GBM cells isolated from human tumors into the brains of severely immuno-deficient mice, or of mouse GBM cells into the brains of congenic host mice. Assays for tumor growth are then performed following sufficient time to allow GSCs among the injected GBM cells to give rise to new tumors-typically several weeks or months. Hence, existing assays do not allow examination of the important pathological process of tumor initiation from single GSCs in vivo. Consequently, essential insights into the specific roles of GSCs and their interaction with the vascular endothelial cells in the early stages of tumor initiation are lacking. Such insights are critical for developing novel therapeutic strategies for glioma, and will have great implications for preventing glioma relapse in patients. Here we have adapted the PoRTS cranial window procedure 16and in vivo two-photon microscopy to allow visualization of tumor initiation from injected GBM cells in the brain of a live mouse. Our technique will pave the way for future efforts to elucidate the key signaling mechanisms between GSCs and vascular endothelial cells during glioma initiation.

Imaging Glioma Initiation In Vivo Through a Polished and Reinforced Thin-skull Cranial Window

By combining a polished and reinforced thin-skull (PoRTS) cranial window and glioblastoma (GBM) cell injection, we can observe glioma initiation and growth from injected GBM cells in the brain of a live mouse longitudinally.

Imágenes Glioma Iniciación<em&gt; En Vivo</em&gt; A través de un pulido y reforzado fina calavera ventana craneal

Glioma is the one of the most lethal forms of human cancer. The most effective glioma therapy to date-surgery followed by radiation treatment-offers ...

A Novel Application of Musculoskeletal Ultrasound Imaging

Ultrasound is an attractive modality for imaging muscle and tendon motion during dynamic tasks and can provide a complementary methodological approach for biomechanical studies in a clinical or laboratory setting. Towards this goal, methods for quantification of muscle kinematics from ultrasound imagery are being developed based on image processing. The temporal resolution of these methods is typically not sufficient for highly dynamic tasks, such as drop-landing. We propose a new approach that utilizes a Doppler method for quantifying muscle kinematics. We have developed a novel vector tissue Doppler imaging (vTDI) technique that can be used to measure musculoskeletal contraction velocity, strain and strain rate with sub-millisecond temporal resolution during dynamic activities using ultrasound. The goal of this preliminary study was to investigate the repeatability and potential applicability of the vTDI technique in measuring musculoskeletal velocities during a drop-landing task, in healthy subjects. The vTDI measurements can be performed concurrently with other biomechanical techniques, such as 3D motion capture for joint kinematics and kinetics, electromyography for timing of muscle activation and force plates for ground reaction force. Integration of these complementary techniques could lead to a better understanding of dynamic muscle function and dysfunction underlying the pathogenesis and pathophysiology of musculoskeletal disorders.

We describe a new ultrasound-based vector tissue Doppler imaging technique to measure muscle contraction velocity, strain and strain rate with sub-millisecond temporal resolution during dynamic activities. This approach provides complementary measurements of dynamic muscle function and could lead to a better understanding of mechanisms underlying musculoskeletal disorders.

Una aplicación Novel of Musculoskeletal Ultrasound Imaging

Ultrasound is an attractive modality for imaging muscle and tendon  motion during dynamic tasks and can provide a complementary methodological  approach ...

Diagnostic Ultrasound Imaging of Mouse Diaphragm Function

Function analysis of rodent respiratory skeletal muscles, particularly the diaphragm, is commonly performed by isolating muscle strips using invasive surgical procedures. Although this is an effective method of assessing in vitro diaphragm activity, it involves non-survival surgery. The application of non-invasive ultrasound imaging as an in vivo procedure is beneficial since it not only reduces the number of animals sacrificed, but is also suitable for monitoring disease progression in live mice. Thus, our ultrasound imaging method may likely assist in the development of novel therapies that alleviate muscle injury induced by various respiratory diseases. Particularly, in clinical diagnoses of obstructive lung diseases, ultrasound imaging has the potential to be used in conjunction with other standard tests to detect the early onset of diaphragm muscle fatigue. In the current protocol, we describe how to accurately evaluate diaphragm contractility in a mouse model using a diagnostic ultrasound imaging technique.

Diagnostic ultrasound imaging has proven to be effective in diagnosing various respiratory diseases in human and animal subjects. We demonstrate a comprehensive ultrasound protocol utilized by Dr. Zuo's lab to analyze diaphragm kinetics specifically in mouse models. This is also a non-invasive research technique which can provide quantitative information on mouse respiratory muscle function.

Ultrasonido Diagnóstico por la Imagen de la función del ratón del diafragma

Function analysis of rodent respiratory skeletal muscles, particularly the diaphragm, is commonly performed by isolating muscle strips using invasive ...

High-frequency Ultrasound Imaging of Mouse Cervical Lymph Nodes

High-frequency ultrasound (HFUS) is widely employed as a non-invasive method for imaging internal anatomic structures in experimental small animal systems. HFUS has the ability to detect structures as small as 30 &#181;m, a property that has been utilized for visualizing superficial lymph nodes in rodents in brightness (B)-mode. Combining power Doppler with B-mode imaging allows for measuring circulatory blood flow within lymph nodes and other organs. While HFUS has been utilized for lymph node imaging in a number of mouse&#160; model systems, a detailed protocol describing HFUS imaging and characterization of the cervical lymph nodes in mice has not been reported. Here, we show that HFUS can be adapted to detect and characterize cervical lymph nodes in mice. Combined B-mode and power Doppler imaging can be used to detect increases in blood flow in immunologically-enlarged cervical nodes. We also describe the use of B-mode imaging to conduct fine needle biopsies of cervical lymph nodes to retrieve lymph tissue for histological&#160; analysis. Finally, software-aided steps are described to calculate changes in lymph node volume and to visualize changes in lymph node morphology following image reconstruction. The ability to visually monitor changes in cervical lymph node biology over time provides a simple and powerful technique for the non-invasive monitoring of cervical lymph node alterations in preclinical mouse models of oral cavity disease.

This protocol describes the application of high-frequency ultrasound (HFUS) for imaging mouse cervical lymph nodes. This technique optimizes visualization and quantification of cervical lymph node morphology, volume and blood flow. Image-guided biopsy of cervical lymph nodes and processing of lymph tissue for histological evaluation is also demonstrated.

Alta frecuencia de imágenes por ultrasonido del mouse ganglios linfáticos cervicales

High-frequency ultrasound (HFUS) is widely employed as a non-invasive method for imaging internal anatomic structures in experimental small animal ...

Transcutaneous Microcirculatory Imaging in Preterm Neonates

Microcirculatory imaging (MI) is a relatively new research tool mainly used in the intensive care setting. MI provides a clear view of the smallest capillaries, arterioles and venules. The magnifying effect visualizes the flow pattern of erythrocytes through these vessels.
It's non-invasive character makes it suitable to apply in (preterm) neonates, even in cardiorespiratory unstable patients. In adults and children, MI is mainly performed sublingually, but this is not possible in preterm infants as these cannot cooperate and the size of the probe is problematic. In preterm infants, MI is therefore performed transcutaneously. Their thin skin makes it possible to obtain high quality images of peripheral microcirculation. 
In this manuscript we will demonstrate the method of transcutaneous MI in preterm infants. We will focus on the different techniques and provide tips to optimize image quality. The highlights of software settings, safety and offline analysis are also addressed.

Microcirculatory imaging (MI) is used to monitor peripheral perfusion in critically ill or preterm neonates. This manuscript and video demonstrates the optimal approach for obtaining high-quality images.

Transcutánea microcirculatoria Imaging en neonatos prematuros

Microcirculatory imaging (MI) is a relatively new research tool mainly used in the intensive care setting. MI provides a clear view of the smallest ...

Ultrasound Imaging of the Thoracic and Abdominal Aorta in Mice to Determine Aneurysm Dimensions

Contemporary high-resolution ultrasound instruments have sufficient resolution to facilitate the measurement of mouse aortas. These instruments have been widely used to measure aortic dimensions in mouse models of aortic aneurysms. Aortic aneurysms are defined as permanent dilations of the aorta, which occur most frequently in the ascending and abdominal regions. Sequential measurements of aortic dimensions by ultrasound are the principal approach for assessing the development and progression of aortic aneurysms in vivo. Although many reported studies used ultrasound imaging to measure aortic diameters as a primary endpoint, there are confounding factors, such as probe position and cardiac cycle, that may impact the accuracy of data acquisition, analysis, and interpretation. The purpose of this protocol is to provide a practical guide on the use of ultrasound to measure the aortic diameter in a reliable and reproducible manner. This protocol introduces the preparation of mice and instruments, the acquisition of appropriate ultrasound images, and data analysis.

Ultrasound imaging has become a common modality to determine the luminal dimensions of thoracic and abdominal aortic aneurysms in mice. This protocol describes the procedure to acquire reliable and reproducible two-dimensional ultrasound images of the ascending and abdominal aorta in mice.

Imágenes por ultrasonido de la Aorta torácica y Abdominal en ratones para determinar dimensiones de aneurisma

Contemporary high-resolution ultrasound instruments have sufficient resolution to facilitate the measurement of mouse aortas. These instruments have been ...