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 segue as orientações do comitê de ética em pesquisa com seres humanos local. 1. Considerações Gerais NOTA: considerações gerais sobre equipamentos, armazenamento e segurança de dados são abordados na discussão. <ol><li> Obter imagens usando uma alta resolução, em tempo real, máquina de ultra-som 2D móvel com vários transdutores com uma banda de frequências (ver discussão). Normalmente, obter imagens de boa qualidade utilizando uma sonda com uma frequência de 7,5 a 8,5 MHz. </li></ol> 2. Preparação do Exame CUS <ol><li> Marcar o exame CUS de modo que ele não coincida com outros processos, tais como a amostragem de sangue. </li><li> Certifique-se que um profissional de saúde ou um pai está disponível para apoiar e / ou o conforto do recém-nascido durante o exame, o uso de estratégias, tais como os de acordo com os princípios do Programa de Avaliação 6 Newborn Individualizado Developmental Cuidados e &lt;/ Sup&gt;. </li></ol> 3. Análise Através Anterior Fontanel <ol><li> Instale a máquina de ultra-som ao longo da incubadora ou berço. </li><li> Aplicar gel transdutor da sonda para garantir um bom contacto entre a sonda e a pele. Considere aquecendo o gel antes da utilização. </li><li> Comece a imagem através da fontanela anterior com uma sonda convexa em modo-B. Coloque a sonda no meio do fontanel com o marcador sobre a sonda virou-se para o lado direito do neonato. O lado esquerdo do cérebro serão mostrados no lado direito do monitor. NOTA: Geração através da fontanela anterior pode ser realizado com o recém-nascido, em qualquer posição 3. Para fins de pesquisa, pode ser necessário para lutar por uma posição da cabeça standard. <ol><li> Grava imagens em pelo menos cinco coronal e sagital cinco. Na primeira imagem ajustar a profundidade, as configurações de ganho de compensação de ganho e de tempo para produzir uma imagem de encher o sector, que contém os contornos cranianos, eviteing imagens muito claras ou escuras e com o objetivo de um equilíbrio entre os reflexos das estruturas vizinhas e mais profundas. </li><li> Planos coronal NOTA: Tente obter imagens perfeitamente simétricas. Quando houver suspeita de lesões próximas da convexidade do lobo frontal, considere a gravação cortes coronais oblíquos específicos, de modo que um hemisfério é exibido em melhor detalhe (Figura 1). <ol><li> Para a imagem pré-frontal coronal, ângulo para a frente da sonda para visualizar os lobos frontais, anterior para os cornos frontais dos ventrículos laterais ao nível dos sulcos olfactiva. </li><li> Para a imagem coronal ao nível de Monro, ângulo da sonda para visualizar a seção anterior coronal ao choroidea Tela para descrever os cornos frontais dos ventrículos laterais, cavum do septo pelúcido, corpo caloso, cinguli sulco. Nota da ecogenicidade de partes do gânglio basal. </li><li> Para a imagem coronal ao nível do tálamo, o ângulo para trás sonda para identificar o tardefissuras ral, choroidea Tela no teto do terceiro ventrículo e lobos temporais. Note-se a ecogenicidade do tálamo (núcleos especialmente ventrolateral) em relação aos gânglios basais. Note-se que lesões na rede pulvinar podem ser visualizados numa secção coronal adicional em frente das aurículas. </li><li> Para a imagem coronal ao nível dos átrios, visualizar os ventrículos laterais ao nível do plexo coróide. Identificar os lobos temporais e hemisférios do cerebelo. Nota da ecogenicidade da substância branca periventricular comparação com plexo coróide. Compare radiação óptica com as áreas hiperecoides acima do normal e laterais para os átrios em recém-nascidos prematuros. </li><li> Para a imagem parieto-occipital coronal, o ângulo de sonda para trás para o nível do sulco parieto-occipital para identificar os lobos parietal e occipital. </li></ol></li><li> Sagital <ol><li> Girar a sonda de 90 ° com o marcador sobre a sonda virada para a face do neonato. The parte anterior do cérebro será exibida no lado esquerdo do monitor. Registam-se as imagens ao nível das seguintes estruturas (Figura 2). </li><li> Para a imagem sagital mediano, visualizar o corpo caloso, cavum do septo pelúcido (CSP), terceiro e quarto ventrículo, vermis, cisterna magna, ponte e mesencéfalo. Observe a presença de cavum vergae e cavum Veli interpositi 7. </li><li> Para a imagem parasagittal através de um ovóide gangliotalâmicas (por exemplo, à direita), o ângulo da sonda para os lados para uma visão parasagittal através do ventrículo lateral. Identificar o plexo coróide e observe a ecogenicidade do tálamo e gânglios basais. O lado digitalizado para aviões parassagitais deve ser devidamente indicada, com ferramentas de texto. </li><li> Para o parasagittal imagem insular, o ângulo da sonda ainda mais lateral através da ínsula. Identificar a fissura lateral e frontal-, temporal-, parietal- e occipital. </li><li> Imagens parassagitais repita para tele lado contralateral (isto é, à esquerda). </li></ol></li><li> Doppler colorido <ol><li> Continue imagens através da fontanela anterior com uma sonda convexa usando Doppler colorido. Considere avaliar as velocidades de fluxo nas artérias cerebrais e veias e obtenção de índices derivados. NOTA: índice de resistência (IR) é definida como velocidade sistólica de pico - velocidade sistólica velocidade diastólica final / pico. RI é independente do ângulo, os valores de velocidade absoluta não são 8-10. RI não é semelhante em artérias de calibre diferente. Medições em série são úteis apenas se for realizada no mesmo local exacto do mesmo recipiente. </li></ol></li><li> Grava imagens em planos coronal dos seguintes vasos (Figura 3): <ol><li> Visualizar as cavidades transversais a nível do cerebelo. Se apenas um ou nenhum dos seios transverso é visualizado, tente diminuir a frequência de repetição do pulso (PRF). Se, ainda assim, apenas um ou nenhum dos seios transversospode ser identificado através da fontanela anterior, use uma sonda linear de alta freqüência para a visualização através da fontanela mastóide (ver secção 4.4.2). </li><li> Visualize o círculo de Willis com as artérias carótidas internas, artérias cerebrais médias e anterior artérias cerebrais ao nível dos cornos frontais dos ventrículos laterais. Distinguir as artérias cerebrais anteriores esquerdo e direito é muitas vezes um desafio, mas é geralmente desnecessário. Identificar o candelabro estriado das artérias. </li><li> Ângulo da sonda para trás para visualizar a artéria basilar com veias jugulares adjacentes. </li><li> Angle ainda mais para trás, para visualizar as veias cerebrais e thalamostriate internos. </li></ol></li><li> Gravar uma imagem no plano sagital de uma artéria cerebral anterior (Figura 4). Avaliar a velocidade do fluxo e RI em uma parte específica deste navio (geralmente abaixo do joelho do corpo caloso). Perto das velocidades da linha média na veia cerebral interna pode serfacilmente medido. </li><li> Usando uma sonda linear de alta freqüência em um plano coronal na fontanela anterior, identificar o seio sagital superior. Se isto falhar, reduzir a quantidade de pressão aplicada com a sonda para o fontanel. NOTA: A sonda linear pode ser usado para visualização detalhada das estruturas superficiais (meninges, espaço aracnóide e subdural, córtex). Vasos são tangenciais no espaço subaracnóideo. Idealmente, o Doppler como descrito nas etapas anteriores serão realizados durante o primeiro exame CUS do neonato. Durante o exame follow-up de algumas das etapas podem ser ignorados. Em caso de suspeita de imagem trombose venosa cerebral Doppler conforme descrito nas etapas 3.3.5.1, 3.3.7 e 4.4.2 devem ser realizados. </li></ol></li></ol> 4. Exame Através Alternativa Acústica do Windows <ol><li> Em seguida, prosseguir a análise através de janelas acústicas alternativas. </li><li> Considere a gravação de imagens através do lambdoid (posterior) Fontanel usando uma sonda convexa (Figura 5). A fontanela posterior está localizado na junção das suturas sagital e lambdóide 3,11. Imagem através da fontanela posterior, colocando o recém-nascido em decúbito lateral. NOTA: Em muitos prematuros imagens satisfatórias também pode ser obtido através do aspecto posterior da sutura sagital com a criança numa posição supina 3. <ol><li> Posicione a sonda no meio da fontanela posterior para uma visão sagital. Ângulo da sonda um pouco fora da linha média para identificar o corpo do ventrículo lateral e seu chifre occipital. Vire a sonda cerca de 90 ° para obter uma visão coronal. Identificar os cornos occipitais dos ventrículos laterais. </li></ol></li><li> Considere gravação de imagens, através da janela lateral (temporal) utilizando uma sonda convexa ou linear acima da orelha (Figura 6). <ol><li> Se necessário, obter imagens através da janela lateral, para permitir umavisão detalhada do tronco cerebral 12. Colocar a sonda na horizontal e ligeiramente acima na frente do ouvido. Mover a sonda até que os pedúnculos cerebrais são visualizados. NOTA: Outras estruturas que podem ser identificados são o terceiro ventrículo, o aqueduto e lobos temporais. Usando cores Doppler, o círculo de Willis pode ser visualizada. </li></ol></li><li> Gravar imagens através da fontanela mastóide (Figura 7). A fontanela mastóide está localizado atrás da orelha, na junção do temporal, parietal e occipital ossos 4. Imagem através da fontanela mastóide, colocando o recém-nascido em uma posição de decúbito lateral 3. NOTA: Em nossa experiência, recém-nascidos muitas vezes mostram sinais de desconforto quando as imagens através da fontanela mastóide são obtidos. Portanto, seria melhor fazer isso depois de imagens através da fontanela anterior e outras janelas acústicas. Nós supomos que este desconforto pode ser causado por o mecanismo de resposta auditiva de pulses de energia de radiofreqüência 13. <ol><li> Imagem através mastóide fontanela usando uma sonda convexa. Coloque a sonda paralela ao ouvido para obter uma visão coronal. Varrer a sonda para trás e para identificar os hemisférios, vermis cerebelar, terceiro e quarto ventrículo, ponte e cisterna magna. Em pequenos prematuros hemisfério cerebelar contralateral pode ser bem representado. </li><li> Imagem através mastóide fontanela usando uma sonda linear. Se (um dos) os seios transversais não pode ser identificado através da fontanela anterior, use uma sonda linear de alta freqüência para a visualização através da fontanela mastóide. Coloque a sonda paralela ao lóbulo da orelha para obter uma visão coronal. <ol><li> Identificar o hemisfério cerebelar e quarto ventrículo. Usando cores Doppler, identificar o transverso e sigmóide, sinus tentorial e veias emissárias. </li></ol></li></ol></li><li> Considere visualização adicional da fossa posterior através do forame magno 14. </li></ol>

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The authors have nothing to disclose.

Nós descrevemos e demonstrar uma abordagem state-of-the-art para CUS Doppler neonatal. Em mãos experientes, esta é uma excelente ferramenta para o seguro, cabeceira de série de imagens do cérebro neonatal. Em muitos UTIN as possibilidades descritas não são plenamente utilizados. Adicionando Doppler permite a triagem da permeabilidade das artérias intracranianas e veias. Velocidades de fluxo pode ser avaliada e índices obtidos. CUS Doppler permite a detecção de trombose venosa cerebral em um estágio inicial n...

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.

Exemplos de imagem feita de acordo com o protocolo descrito são apresentados nas Figuras 1-7. As imagens devem ser interpretados com cautela por um observador experiente. Imagem simétrica é necessário para interpretação das imagens coronais adequada feitas através da fontanela anterior (Figura 1). Qualquer lesão suspeita deve ser visualizada em ambos um e (mid) plano sagital coronal ou através da visualização através de uma janela acústica diferente da fontanela anterior. U...

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 da Arte Cranial Ecografia em recém-nascidos

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 visualizações.  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. 

Vídeo: 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.

Dual-mode de imagem de oxigenação dos tecidos cutâneos e Função Vascular

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

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.

Ecocardiografia murino e ultra-sonografia

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.

Guiada por MRI rompimento da barreira hemato-encefálica utilizando Ultrassom Focalizado transcraniana em um modelo de rato

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.

Multi-modal de imagem de angiogênese em um modelo de rato nu de Câncer de mama utilizando a ressonância magnética, tomografia computadorizada volumétrica e ultra-som

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.

Imagem Glioma Iniciação<em&gt; Em Vivo</em&gt; Através de um polido e reforçado Janela crânio-Thin craniana

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.

A Aplicação da novela da Musculoskeletal Ecografia

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.

Diagnóstico Ecografia de função de mouse 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.

De alta frequência Ecografia de mouse linfonodos cervicais

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.

Transcutânea Microcirculatory Imaging no recém-nascidos pré-termo

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.

Ecografia da Aorta torácica e Abdominal em ratos para determinar as dimensões de aneurisma

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