We thank J. Hagoort for reading and correcting the manuscript.

Este protocolo segue as diretrizes do comitê de ética em pesquisa humanos local. 1. Preparação <ol><li> Marcar a medição da microcirculação de modo que ele não coincida com outros processos, tais como a amostragem de sangue. Em neonatos a termo que é melhor executada após a alimentação. Isto evita a agitação e vai facilitar a medição. </li><li> Certifique-se de que uma enfermeira ou um pai atende a apoiar e confortar o recém-nascido durante o exame, usando os princípios do Programa de Avaliação do recém-nascido 12 Individualizado Desenvolvimento e Cuidados. Nota: Embora as medições podem ser realizadas por uma única pessoa, é altamente recomendável para ter uma segunda pessoa auxiliar. Um prende a câmera e está focada no recém-nascido, enquanto o outro opera o computador e software. Em nossa experiência, isso resulta em imagens de alta qualidade e uma duração mais curta do procedimento. </li><li> Se a condição clínica dos neonatos licenças, coloque oneonato em posição supina. Imagem da microcirculação pode ser realizada em decúbito ventral, mas isso requer mais habilidade e paciência. </li><li> Verifique se a temperatura do corpo do recém-nascido prematuro está dentro do alcance adequado (36,5 - 37,5 graus Celsius). </li></ol> 2. Procedimento <ol><li> Instale o dispositivo ao longo da incubadora. Certifique-se da incubadora é a altura certa. </li><li> Coloque a tampa descartável na câmera. </li><li> Aplicar gel, óleo ou solução salina na ponta da sonda; isso vai ajudar a suavizar o contato entre a sonda ea pele. </li><li> Coloque a câmera no lado ventromedial do recém braço. Para evitar que o foco-artefactos, certifique-se a sonda é perpendicular à pele. Isto pode exigir o reposicionamento do braço lactentes. Nota: O lado ventromedial do braço é o local principal para medir a microcirculação cutânea. Este local tem pouco cabelo lanugo e é, portanto, menos propenso a artefatos. Ele é mais facilmente alcançadoSe o paciente é posicionado em posição supina. </li><li> Para minimizar o comprimento total do processo, ganhar tempo, encontrando a profundidade óptima de foco (Figura 3), enquanto procura o local com o menor número de produtos manufacturados. Nota: Profundidade do foco depende principalmente da idade pós-natal, em vez de idade gestacional. A profundidade média de foco na primeira semana de vida é de 0 - 80 um. Daqui por diante, devido ao amadurecimento da pele, a profundidade de foco aumenta rapidamente com valores médios de 80 - 200 um entre 1 - 4 semanas de idade pós-natal (Figura 2). No prazo nascido recém-nascidos a profundidade média de foco é 80-160 mm no nascimento. </li><li> Estabilizar a sonda para evitar artefatos de movimento. Para fazê-lo, o cotovelo descansar na janela da incubadora e o pulso ao lado do neonato. Alternativamente, posicionar a sonda ao lado do recém-nascido em um travesseiro. </li><li> Evite artefatos de pressão, permitindo que a câmara só tem a menor contato com a pele. Artefatos de pressão pode ser recognized durante captura de imagem se há fluxo de vai-e-vem em vasos ou se grandes vasos são não-perfundidos enquanto houver bom fluxo em pequenas embarcações. Além disso, se o padrão de fluxo é idêntica em toda a tela inteira, cuidado com os artefatos de pressão. </li><li> Gravar vídeos com uma duração mínima de 5 segundos. </li><li> Depois de uma captação bem sucedida, mova a câmera para outro local na parte superior do braço. Nota: Recomenda-se capturar um total de 5 - 10 vídeos no 3 - 5 locais diferentes, como alguns artefactos só são reconhecidos na análise off-line, o que significa que o vídeo em questão não é utilizável para análise. </li><li> Suavemente remover o gel, óleo ou solução salina a partir da pele com um pequeno gaze. </li></ol> 3. Análise offline <ol><li> Recortar o vídeo, se há um movimento significativo que impede a análise. Vá para a seção &#39;Ferramentas&#39; e use o botão &#39;Editor&#39;. Selecione o intervalo de quadro elegíveis para análise e clique no &#39;Vídeo da colheita&#39; button. Nota: Os vídeos são aceitáveis ​​se o movimento está dentro de ½ do campo de visão 13. </li><li> Selecione o vídeo recortada e estabilizá-lo. Vá para a seção &#39;Ferramentas&#39; e use o botão &#39;Análise&#39;. Clique no botão &quot;Estabilizar&quot;. Nota: Todos os filmes devem ser estabilizados antes da análise automática pode ser executada. </li><li> Selecione o vídeo estabilizado. Vá para a seção &quot;análise&quot; e clique no botão &#39;Detect&#39;. Certifique-se destacam as opções &#39;Capilares »e« navios ». </li><li> Após a detecção (Figura 4), ​​clique no botão &#39;De Backer&#39; &#39;CNA &quot;ou para um relatório completo da microcirculação. Este relatório inclui os parâmetros mais usados ​​de resultados como a densidade total do vaso (TVD), densidade de vasos perfundidos (PVD) e proporção de vasos perfundidos (PPV). Nota: Como alternativa, os vídeos podem ser exportados offline para ser analisada manualmente. Esta opção pode ser encontrada nas &#39;Ferramentas&#39; seção. Seleccione a opção &#39;Exportar&#39; e clique no botão &#39;AVA exportação ». </li></ol>

<ol>
	<li>Groner, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthogonal+polarization+spectral+imaging:+a+new+method+for+study+of+the+microcirculation.">Orthogonal polarization spectral imaging: a new method for study of the microcirculation.</a> Nat Med. 5 (10), 1209-1212 (1999).</li><li>Goedhart, P. T., Khalilzada, M., Bezemer, R., Merza, J., Ince, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sidestream+Dark+Field+(SDF)+imaging:+a+novel+stroboscopic+LED+ring-based+imaging+modality+for+clinical+assessment+of+the+microcirculation.">Sidestream Dark Field (SDF) imaging: a novel stroboscopic LED ring-based imaging modality for clinical assessment of the microcirculation.</a> Opt Express. 15 (23), 15101-15114 (2007).</li><li>Sherman, H., Klausner, S., Cook, W. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Incident+dark-field+illumination:+a+new+method+for+microcirculatory+study.">Incident dark-field illumination: a new method for microcirculatory study.</a> Angiology. 22 (5), 295-303 (1971).</li><li>Trzeciak, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+microcirculatory+perfusion+derangements+in+patients+with+severe+sepsis+and+septic+shock:+relationship+to+hemodynamics,+oxygen+transport,+and+survival.">Early microcirculatory perfusion derangements in patients with severe sepsis and septic shock: relationship to hemodynamics, oxygen transport, and survival.</a> Ann Emerg Med. 49 (1), 88-98 (2007).</li><li>Sakr, Y., Dubois, M. J., De Backer, D., Creteur, J., Vincent, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Persistent+microcirculatory+alterations+are+associated+with+organ+failure+and+death+in+patients+with+septic+shock.">Persistent microcirculatory alterations are associated with organ failure and death in patients with septic shock.</a> Crit Care Med. 32 (9), 1825-1831 (2004).</li><li>De Backer, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microcirculatory+alterations+in+patients+with+severe+sepsis:+impact+of+time+of+assessment+and+relationship+with+outcome.">Microcirculatory alterations in patients with severe sepsis: impact of time of assessment and relationship with outcome.</a> Crit Care Med. 41 (3), 791-799 (2013).</li><li>Buijs, E. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+microcirculatory+impairment+during+therapeutic+hypothermia+is+associated+with+poor+outcome+in+post-cardiac+arrest+children:+A+prospective+observational+cohort+study.">Early microcirculatory impairment during therapeutic hypothermia is associated with poor outcome in post-cardiac arrest children: A prospective observational cohort study.</a> Resuscitation. , (2013).</li><li>Genzel-Boroviczeny, O., Christ, F., Glas, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood+transfusion+increases+functional+capillary+density+in+the+skin+of+anemic+preterm+infants.">Blood transfusion increases functional capillary density in the skin of anemic preterm infants.</a> Pediatr Res. 56 (5), 751-755 (2004).</li><li>Ergenekon, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Peripheral+microcirculation+is+affected+during+therapeutic+hypothermia+in+newborns.">Peripheral microcirculation is affected during therapeutic hypothermia in newborns.</a> Arch Dis Child Fetal Neonatal Ed. 98 (2), F155-F157 (2013).</li><li>Schwepcke, A., Weber, F. D., Mormanova, Z., Cepissak, B., Genzel-Boroviczeny, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microcirculatory+mechanisms+in+postnatal+hypotension+affecting+premature+infants.">Microcirculatory mechanisms in postnatal hypotension affecting premature infants.</a> Pediatr Res. , (2013).</li><li>van Elteren, H. A., Ince, C., Tibboel, D., Reiss, I. K., de Jonge, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cutaneous+microcirculation+in+preterm+neonates:+comparison+between+sidestream+dark+field+(SDF)+and+incident+dark+field+(IDF)+imaging.">Cutaneous microcirculation in preterm neonates: comparison between sidestream dark field (SDF) and incident dark field (IDF) imaging.</a> J Clin Monit Comput. , (2015).</li><li>Als, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Individualized+Behavioral+and+Environmental+Care+for+the+Very-Low-Birth-Weight+Preterm+Infant+at+High-Risk+for+Bronchopulmonary+Dysplasia+-+Neonatal+Intensive-Care+Unit+and+Developmental+Outcome.">Individualized Behavioral and Environmental Care for the Very-Low-Birth-Weight Preterm Infant at High-Risk for Bronchopulmonary Dysplasia - Neonatal Intensive-Care Unit and Developmental Outcome.</a> Pediatrics. 78 (6), 1123-1132 (1986).</li><li>Massey, M. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+microcirculation+image+quality+score:+development+and+preliminary+evaluation+of+a+proposed+approach+to+grading+quality+of+image+acquisition+for+bedside+videomicroscopy.">The microcirculation image quality score: development and preliminary evaluation of a proposed approach to grading quality of image acquisition for bedside videomicroscopy.</a> J Crit Care. 28 (6), 913-917 (2013).</li><li>Weidlich, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Changes+in+microcirculation+as+early+markers+for+infection+in+preterm+infants--an+observational+prospective+study.">Changes in microcirculation as early markers for infection in preterm infants--an observational prospective study.</a> Pediatr Res. 66 (4), 461-465 (2009).</li><li>De Backer, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=How+to+evaluate+the+microcirculation:+report+of+a+round+table+conference.">How to evaluate the microcirculation: report of a round table conference.</a> Crit Care. 11 (5), R101 (2007).</li><li>Sallisalmi, M., Oksala, N., Pettila, V., Tenhunen, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+sublingual+microcirculatory+blood+flow+in+the+critically+ill.">Evaluation of sublingual microcirculatory blood flow in the critically ill.</a> Acta Anaesthesiol Scand. 56 (3), 298-306 (2012).</li><li>van den Berg, V. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reproducibility+of+microvascular+vessel+density+analysis+in+Sidestream+dark-field-derived+images+of+healthy+term+newborns.">Reproducibility of microvascular vessel density analysis in Sidestream dark-field-derived images of healthy term newborns.</a> Microcirculation. 22 (1), 37-43 (2015).</li></ol>

Os autores não têm nada a revelar.

Nesse artigo, procuramos descrever e demonstrar a abordagem para geração de imagens da microcirculação transcutânea em recém-nascidos prematuros. Visualizando esse método irá ajudar os pesquisadores a dois dos maiores desafios na investigação: reprodutibilidade e o tempo eo trabalho natureza intensiva de aprender novas técnicas. Esta técnica pode proporcionar informação útil de microcirculação periférica em prematuros de forma não-invasiva. Medições em série pode ajudar os médicos a avaliar os efe...

Hemodynamic diagnostics in critically ill preterm neonates has always been difficult. Most diagnostic tools used in adults cannot be applied in these tiny preterm infants; and then there is a problem of the sensitivity of the outcome parameters. But most of all, these infants are so vulnerable, that the risks of diagnostic procedures do not outweigh the benefits. As a result, in the field of neonatology, hemodynamics has been neglected and therefore there is a lack of knowledge on this topic.
An interesting option for handling these problems might be visualizing the microcirculation. The introduction of handheld microscopes in the late 1990s made it possible to visualize the microcirculation in a non-invasive manner. Three generations of devices have been introduced: Orthogonal Polarization Spectral (OPS) imaging1, Sidestream Dark Field (SDF) imaging2, and Incident Dark Field (IDF) imaging3. They all use more or less the same technique in which green light with a specific wavelength (548nm) stroboscopic illuminates the microcirculation. The green light is absorbed by oxy- and deoxyhemoglobin and mostly reflected by the surrounding tissue. This property of green light therefore creates visible contrast. The reflected light passes a magnification lens and is projected on a camera sensor. Hereby it is possible to visualize the flowing red blood cells at a depth of approximately one millimeter of mucosal tissue or directly at solid organs. 
Over the past 15 years, the microcirculation has been mainly studied in adults, especially in patients with septic shock4-6. These observational studies found that persistent microcirculatory alterations were associated with organ failure and mortality. This observation cannot be extrapolated directly to (preterm) infants however, as in the adults the microcirculation was measured sublingually. High quality images of the sublingual microcirculation cannot be obtained in preterm infants because they are unable to cooperate. In term infants the buccal microcirculation has been the area of interest7. Fortunately, in preterm infants the thin skin allows transcutaneous microcirculatory imaging. This approach has been applied in neonatal studies focusing on blood transfusion8, therapeutic hypothermia9 and hypotension10.
In this manuscript we present our protocol for transcutaneous microcirculatory imaging using Incident Dark Field imaging in preterm neonates. We will focus on different strategies to acquire the highest quality images. Technical details and differences between the SDF and IDF devices can be found elsewhere11.

<table>
	<tbody>
		<tr>
			<td>Cytocam</td>
			<td>Braedius</td>
			<td>http://www.braedius.com/magnoliaPublic/braedius/products.html</td>
			<td>Other well known handheld microscopes to visualize the microcirculation are MicroScan (Microvision Medical) using SDF technique or the CytoScan (CytoMetrics) using OPS technique</td>
		</tr>
		<tr>
			<td>Disposable Lens Cover</td>
			<td>Glycocheck</td>
			<td>http://www.glycocheck.com/lenscovers.php</td>
			<td></td>
		</tr>
		<tr>
			<td>CCTools</td>
			<td>Braedius</td>
			<td>http://www.braedius.com/magnoliaPublic/braedius/products.html</td>
			<td>Another well known offline analysis programme is AVA (Microvision medical).&#160;</td>
		</tr>
	</tbody>
</table>

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.

As Figuras 1 e 2 mostram representante de imagens fixas de alta qualidade vídeos mi. Estes exemplos demonstram a diferença da espessura da pele em crianças entre os mesmos dia 1 (Figura 1) e no dia 28 (Figura 2) da idade pós-natal. No dia 1, há uma iluminação brilhante, foco adequada sobre os micro vasos e presença mínima de artefatos. No dia 28, é mais difícil encontrar o equilíbrio certo entre foco nas micro vasos e artefat...

Watch this Scientific Journal Video about Transcutaneous Microcirculatory Imaging in Preterm Neonates at JoVE.com

Transcutaneous Microcirculatory Imaging in Preterm Neonates

Transcutânea Microcirculatory Imaging no recém-nascidos pré-termo

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.

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

transcutaneous-microcirculatory-imaging-in-preterm-neonates

Research

JoVE Journal

Medicine

7.6K visualizações.  Erasmus MC-Sophia Children's Hospital. 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.

Vídeo: Transcutaneous Microcirculatory Imaging in Preterm Neonates

The Use of Pharmacological-challenge fMRI in Pre-clinical Research: Application to the 5-HT System

Pharmacological MRI (phMRI) is a new and promising method to study the effects of substances on brain function that can ultimately be used to unravel underlying neurobiological mechanisms behind drug action and neurotransmitter-related disorders, such as depression and ADHD. Like most of the imaging methods (PET, SPECT, CT) it represents a progress in the investigation of brain disorders and the related function of neurotransmitter pathways in a non-invasive way with respect of the overall neuronal connectivity. Moreover it also provides the ideal tool for translation to clinical investigations. MRI, while still behind in molecular imaging strategies compared to PET and SPECT, has the great advantage to have a high spatial resolution and no need for the injection of a contrast-agent or radio-labeled molecules, thereby avoiding the repetitive exposure to ionizing radiations. Functional MRI (fMRI) is extensively used in research and clinical setting, where it is generally combined with a psycho-motor task. phMRI is an adaptation of fMRI enabling the investigation of a specific neurotransmitter system, such as serotonin (5-HT), under physiological or pathological conditions following activation via administration of a specific challenging drug.
The aim of the method described here is to assess brain 5-HT function in free-breathing animals. By challenging the 5-HT system while simultaneously acquiring functional MR images over time, the response of the brain to this challenge can be visualized. Several studies in animals have already demonstrated that drug-induced increases in extracellular levels of e.g. 5-HT (releasing agents, selective re-uptake blockers, etc) evoke region-specific changes in blood oxygenation level dependent (BOLD) MRI signals (signal due to a change of the oxygenated/deoxygenated hemoglobin levels occurring during brain activation through an increase of the blood supply to supply the oxygen and glucose to the demanding neurons) providing an index of neurotransmitter function. It has also been shown that these effects can be reversed by treatments that decrease 5-HT availability16,13,18,7. In adult rats, BOLD signal changes following acute SSRI administration have been described in several 5-HT related brain regions, i.e. cortical areas, hippocampus, hypothalamus and thalamus9,16,15. Stimulation of the 5-HT system and its response to this challenge can be thus used as a measure of its function in both animals and humans2,11.

The goal of this technique is to assess serotonin (5-HT) neurotransmitter function in the live and free-breathing animal with pharmacological magnetic resonance imaging (phMRI) and an intravenous challenge with a selective serotonin reuptake inhibitor (SSRI), fluoxetine.

O Uso de desafio-Farmacológicas fMRI em pesquisa pré-clínica: aplicação ao sistema 5-HT

Pharmacological MRI (phMRI) is a new and promising method to study the effects of substances on brain function that can ultimately be used to unravel ...

Medicina

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 ...

Automated Measurement of Microcirculatory Blood Flow Velocity in Pulmonary Metastases of Rats

Because the lung is a major target organ of metastatic disease, animal models to study the physiology of pulmonary metastases are of great importance. However, very few methods exist to date to investigate lung metastases in a dynamic fashion at the microcirculatory level, due to the difficulty to access the lung with a microscope. Here, an intravital microscopy method is presented to functionally image and quantify the microcirculation of superficial pulmonary metastases in rats, using a closed-chest pulmonary window and automated analysis of blood flow velocity and direction. The utility of this method is demonstrated to measure increases in blood flow velocity in response to pharmacological intervention, and to image the well-known tortuous vasculature of solid tumors. This is the first demonstration of intravital microscopy on pulmonary metastases in a closed-chest model. Because of its minimized invasiveness, as well as due to its relative ease and practicality, this technology has the potential to experience widespread use in laboratories that specialize on pulmonary tumor research.

A method is presented to measure microcirculatory blood flow velocity in pulmonary cancer metastases of the pleural surface in rats in an automated fashion, using closed-chest pulmonary intravital microscopy. This model has potential to be used as a widespread tool to perform physiologic research on pulmonary metastases in rodents.

Automated Medição da microcirculação sanguínea velocidade na Pulmonar Metástases of Rats

Because the lung is a major target organ of metastatic disease, animal models to study the physiology of pulmonary metastases are of great importance. ...

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.

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 ...

Transcutaneous Assessment of Renal Function in Conscious Rodents

Glomerular filtration rate (GFR) is the gold standard to assess overall kidney function. However, traditional methods to evaluate GFR are cumbersome and time-consuming. In addition, serial blood or urine samples are required, with the associated stress for the experimental animals. A recent technique significantly reduces the investment in time and resources, minimizing the invasiveness and the animal stress, but being equally valid as the traditional approaches. The method measures transcutaneously renal function. Using an optical device and the exogenous renal marker fluorescein isothiocyanate (FITC)-sinistrin, this technique is capable of measuring the elimination kinetics of the marker through the skin. With neither blood nor urine samples nor the associated laboratory assays needed, the results of the transcutaneous measurement are almost instantaneously available. The method has been already validated in different species and successfully applied in several models of renal pathology. Moreover, due to its minimally invasive characteristics, it is suitable for sequential measurements within the same animal. Here is provided a detailed protocol to carry out the transcutaneous assessment of renal function in rodents.

Determination of glomerular filtration rate (GFR) is the gold standard to assess overall kidney function. However, traditional procedures to measure this parameter are cumbersome and require a large investment of time. Here we describe a faster and minimally invasive method to determinate GFR transcutaneously.

Avaliação transcutânea da função renal em roedores conscientes

Glomerular filtration rate (GFR) is the gold standard to assess overall kidney function. However, traditional methods to evaluate GFR are cumbersome and ...

Quantitative Visualization of Leukocyte Infiltrate in a Murine Model of Fulminant Myocarditis by Light Sheet Microscopy

Light-sheet fluorescence microscopy (LSFM), in combination with chemical clearing protocols, has become the gold standard for analyzing fluorescently labelled structures in large biological specimens, and is down to cellular resolution. Meanwhile, the constant refinement of underlying protocols and the enhanced availability of specialized commercial systems enable us to investigate the microstructure of whole mouse organs and even allow for the characterization of cellular behavior in various live-cell imaging approaches. Here, we describe a protocol for the spatial whole-mount visualization and quantification of the CD45+ leukocyte population in inflamed mouse hearts. The method employs a transgenic mouse strain (CD11c.DTR)that has recently been shown to serve as a robust, inducible model for the study of the development of fulminant fatal myocarditis, characterized by lethal cardiac arrhythmias. This protocol includes myocarditis induction, intravital antibody-mediated cell staining, organ preparation, and LSFM with subsequent computer-assisted image post-processing. Although presented as a highly-adapted method for our particular scientific question, the protocol represents the blueprint of an easily adjustable system that can also target completely different fluorescent structures in other organs and even in other species.

Here, we describe a light-sheet microscopy approach to visualize the cardiac CD45+ leukocyte infiltrate in a murine model of aseptic fulminant myocarditis, which is induced by the intratracheal diphtheria toxin treatment of CD11c.DTR mice.

Visualização Quantitativa de Infiltrado de Leucócitos em um Modelo Murino de Miocardite Fulminante por Microscopia de Folha de Luz

Light-sheet fluorescence microscopy (LSFM), in combination with chemical clearing protocols, has become the gold standard for analyzing fluorescently ...

Application of an Amplitude-integrated EEG Monitor (Cerebral Function Monitor) to Neonates

Amplitude-integrated EEG (aEEG) is an easily accessible technique to monitor the electrocortical activity in preterm and term infants in neonatal intensive care units (NICUs). This method was first used to monitor newborns after asphyxia, providing information about future neurological outcomes. The aEEG is also helpful to select newborns who benefit from cooling. The aEEG monitoring of preterm infants is becoming more widespread, as various studies have shown that neurodevelopmental outcome is related to early aEEG tracings. Here, we demonstrate the application of the aEEG monitoring system and present typical patterns that depend upon gestational age and pathophysiological conditions. Furthermore, we mention pitfalls in the interpretation of the aEEG, as this method requires accurate fixation and localization of the electrodes. Additionally, the raw EEG can be used to detect neonatal seizures or to identify aEEG application problems. In conclusion, aEEG is a safe and generally well-tolerated method for the bedside monitoring of neonatal cerebral function; it can even provide information about long-term outcome.

Application of an Amplitude-integrated EEG Monitor Cerebral Function Monitor to Neonates

Here, we show how to apply amplitude-integrated electroencephalography to monitor cerebral function in neonates.

Aplicação de um Monitor de EEG Amplitude-integrado (Monitor de função Cerebral) para recém-nascidos

Amplitude-integrated EEG (aEEG) is an easily accessible technique to monitor the electrocortical activity in preterm and term infants in neonatal ...

Electrophysiological Measurement of Noxious-evoked Brain Activity in Neonates Using a Flat-tip Probe Coupled to Electroencephalography

Pain is an unpleasant sensory and emotional experience. In non-verbal patients, it is very difficult to measure pain, even with pain assessment tools. Those tools are subjective or determine secondary physiological indicators which also have certain limitations particularly when exploring the effectiveness of analgesia. As cortical processing is essential for pain perception, brain activity measures may provide a useful approach to assess pain in infants. Here we present a method to assess nociception with electrophysiological brain activity recordings optimized for the use in newborn infants. To produce highly standardized and reproducible noxious stimuli we applied mechanical stimulation with a flat-tip probe, e.g., PinPrick, which is not skin-breaking and does not cause behavioral distress. The noxious-evoked potential allows the objective measurement of nociception in non-verbal patients. This method can be used in newborn infants as early as 34 weeks of gestational age. Moreover, it could be applied in different situations such as measuring the efficacy of analgesic or anesthetic drugs.

Measuring pain in non-verbal patients is a challenge. In this study we combine EEG recording with stimulation using a flat-tip probe to detect noxious-evoked brain activity in an objective manner.

Eletrofisiológica medição da atividade cerebral evocados nocivas em neonatos usando uma sonda de Flat-ponta acoplada a Eletroencefalografia

Pain is an unpleasant sensory and emotional experience. In non-verbal patients, it is very difficult to measure pain, even with pain assessment tools. ...

Phase Contrast Magnetic Resonance Imaging in the Rat Common Carotid Artery

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous studies have shown that abnormal blood flow may be associated with systemic vascular risk. Thus, PC-MRI can facilitate the translation of data obtained from animal models of cardiovascular diseases to pertinent clinical investigations. In this report, we describe the procedure for measuring blood flow in the common carotid artery (CCA) of rats using cine-gated PC-MRI and discuss relevant analysis methods. This procedure can be performed in a live, anesthetized animal and does not require euthanasia after the procedure. The proposed scanning parameters yield repeatable measurements for blood flow, indicating excellent reproducibility of the results. The PC-MRI procedure described in this article can be used for pharmacological testing, pathophysiological assessment, and cerebral hemodynamics evaluation.

The overall goal of this procedure is to measure the blood flow in the rat common carotid artery by using noninvasive phase contrast magnetic resonance imaging.

Imagens de ressonância magnética fase contraste na artéria carótida comum de rato

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous ...

Quantitative [18F]-Naf-PET-MRI Analysis for the Evaluation of Dynamic Bone Turnover in a Patient with Facetogenic Low Back Pain

Imaging techniques that reflect dynamic bone turnover may aid in characterizing a wide range of bone pathologies. Bone is a dynamic tissue undergoing continuous remodeling with the competing activity of osteoblasts, which produce the new bone matrix, and osteoclasts, whose function is to eliminate mineralized bone. [18F]-NaF is a positron emission tomography (PET) radiotracer that enables visualization of bone metabolism. [18F]-NaF is chemically absorbed into hydroxyapatite in the bone matrix by osteoblasts and can thus noninvasively detect osteoblastic activity, which is occult to conventional imaging techniques. Kinetic modeling of dynamic [18F]-NaF-PET data provides detailed quantitative measures of bone metabolism. Conventional semi-quantitative PET data, which utilizes standardized uptake values (SUVs) as a measure of radiotracer activity, is referred to as a static technique due to its snapshot of tracer uptake in time.&#160; Kinetic modeling, however, utilizes dynamic image data where tracer levels are continuously acquired providing tracer uptake temporal resolution. From the kinetic modeling of dynamic data, quantitative values like blood flow and metabolic rate (i.e., potentially informative metrics of tracer dynamics) can be extracted, all with respect to the measured activity in the image data. When combined with dual modality PET-MRI, region-specific kinetic data can be correlated with anatomically registered high-resolution structural and pathologic information afforded by MRI. The goal of this methodological manuscript is to outline detailed techniques for performing and analyzing dynamic [18F]-NaF-PET-MRI data. The lumbar facet joint is a common site of degenerative arthritis disease and a common cause for axial low back pain.&#160; Recent studies suggest [18F]-NaF-PET may serve as a useful biomarker of painful facetogenic disease.&#160; The human lumbar facet joint will, therefore, be used as a prototypical region of interest for dynamic [18F]-NaF-PET-MRI analysis in this manuscript.

Quantitative [18F]-Naf-PET-MRI Analysis for the Evaluation of Dynamic Bone Turnover in a Patient with Facetogenic Low Back Pain

Imaging techniques that reflect dynamic bone turnover may aid in characterizing a wide range of bone pathologies. We present detailed methodologies for performing and analyzing dynamic [18F]-NaF-PET-MRI data in a patient with facetogenic low back pain using the lumbar facet joints as a prototypical region of interest.

Análise quantitativa [18F]-NAF-PET-RM para avaliação da rotatividade óssea dinâmica em paciente com dor nas costas Facetogênica

Imaging techniques that reflect dynamic bone turnover may aid in characterizing a wide range of bone pathologies. Bone is a dynamic tissue undergoing ...