Financiamiento proporcionado por el NIMH, NARSAD y la Fundación Gatsby.

Configuración del equipo El primer paso en la dirección y el experimento fMRI es la creación y la comprobación del equipo. Estos pasos se pueden hacer en cualquier orden. Acondicionamiento de la señal - El escáner se encuentra en una sala eléctrica y blindaje magnético. Todas las señales eléctricas que va de la sala de control para la sala del escáner pasar a través de un panel de filtro para eliminar todas las frecuencias que podrían crear un artefacto en la imagen de RM. Airpuff - El dispositivo airpuff aplica un soplo controlado de la presión y la duración fija. Cuando se dirige hacia el ojo, esto es desagradable, pero no traumática. El regulador de presión (WPI neumático PicoPump) a una fuente de aire comprimido, que podría ser un tanque de aire, pero nosotros usamos la casa del aire. La presión se ajusta a 30 psi. El tiempo es ajustado para ofrecer una bocanada 50 ms. La válvula solenoide es controlada por una señal de ordenador. La salida del regulador de presión se entrega al sujeto a través de un tubo de plástico 1/6o pulgadas (Tygon). Es importante que el tubo es demasiado largo, o habrá un retraso entre el solenoide de apertura y el tiempo que el aire comprimido llega al extremo del tubo. Por esta razón, el regulador de presión se encuentra dentro de la sala del escáner y las señales de control se alimentan a través de un panel de filtros para asegurar que no hay artefactos de imagen. Jugo - El dispensador de jugo controla el tamaño y el número de gotas de jugo de entrega a la boca por un tubo de plástico. El dispensador de jugo consta de un depósito, una válvula solenoide controlada por ordenador, y un tubo largo que ofrece el jugo al tema. Todos los componentes electrónicos están fuera de la sala del escáner y por lo tanto no introducir artefactos en la señal de RM. El dispensador de jugo (Instrumentos Crist) se lavaron dos veces por el llenado del embalse y la apertura de la válvula solenoide. A continuación, se llena con la bebida preferida del sujeto. El sistema es probado antes de la digitalización mediante la ejecución de un programa que hace que el equipo para generar una corriente de impulsos de prueba que se traduce en unas gotas de jugo de ser entregados a la materia. El tema verifica que el jugo que fluye. Mordida bar - El movimiento principal crea varios artefactos en la imagen de resonancia magnética funcional. En este experimento, los sujetos reciben airpuffs que puede provocar movimientos reflejos, y el jugo que se requiere para tragar. Para minimizar el movimiento de la cabeza, un bitebar se utiliza para estabilizar la cabeza. El bitebar está conectado a la bobina de la cabeza de RF. Cada materia tiene una boquilla hecha a la medida de un material termoplástico. Oxímetro de pulso - FMRI medidas de oxigenación de la sangre (BOLD). Oxigenación de la sangre en el cerebro pueden ser afectados por la actividad neuronal, sino que también puede verse afectada por los latidos del corazón. Para medir los cambios en la oxigenación en relación con el ciclo cardíaco, se utiliza un oxímetro de pulso para medir la oxigenación de la sangre en la punta de los dedos usando un sensor infrarrojo. El oxímetro de pulso es compatible con RM y su señal de salida se alimenta a través de un panel de filtros a la sala de control donde se digitaliza y almacena en el ordenador. Monitor de gases respiratorios - oxigenación de la sangre también puede verse afectada por la respiración. Medimos la respiración con un monitor de gas respiratorio que mide los niveles de CO2 expirado. RGM es compatible con RM y su señal de salida se alimenta a través de un panel de filtros a la sala de control donde se digitaliza y almacena en el ordenador. La estimulación visual - Un panel de retroproyección se encuentra en un extremo del escáner llevaba cerca de la ubicación de la cabeza respecto. Temas mirar a la pantalla de proyección trasera a través de un espejo montado en la parte superior de la bobina de la cabeza. Las imágenes se proyectan en la pantalla de proyección trasera de un proyector LCD situado fuera de la sala del escáner. El haz de luz del proyector se dirige dentro de la sala del escáner a través de una manguera de blindaje RF. Seguimiento de los ojos - Es importante saber dónde está el sujeto está mirando por varias razones. Movimientos de los ojos afecta a la posición de los estímulos visuales en la retina, que afecta a las respuestas visuales en el cerebro. Movimientos de los ojos también puede ser utilizado como un medio para que el sujeto indica una respuesta conductual. Nosotros utilizamos un método llamado &quot;oculografía video infrarrojo&quot; para realizar un seguimiento de los ojos. Según este método, una cámara de infrarrojos se utiliza para realizar el seguimiento del alumno. El emisor de infrarrojos y la cámara están integrados en un par de anteojos especialmente diseñados. Estas son las gafas mismos que proporcionan un estímulo visual. Los datos de las gafas es procesada por un equipo dedicado que convierte las imágenes del ojo en señales analógicas para la posición de los ojos horizontales y verticales. Preparación tema Documentación y Consentimiento &amp; ndash, Antes de escanear, cada sujeto debe llenar varios formularios en materia de seguridad y consentimiento. Hoja plan experimental - La hoja de plan especifica el protocolo experimental y las secuencias de pulso y el orden en que se ejecutan. Preparación física - Cuando el sujeto está listo para ser escaneado, una de las primeras cosas que hacen es poner en tapones para los oídos para proteger su audición por el ruido de frecuencia alta y escáner de alta. También pusieron en la RM compatible con auriculares para comunicarse tanto con los sujetos en el escáner y ofrecer instrucciones y comentarios durante la sesión de exploración. El tema pone en su boquilla, que incluye 1 / 16 pulgadas para la entrega de las líneas de jugos y vigilancia de los gases respiratorios. El tema se pone las gafas y ajustar MR compatible utiliza para la estimulación visual y el seguimiento de movimientos oculares. El tema se encuentra en el escáner y la bobina de RF se coloca sobre su cabeza. La boquilla (bitebar) está conectado a la bobina de RF para evitar movimientos de la cabeza. El tubo airpuff (tygon 1 / 6 pulgadas) se coloca cerca de un ojo con un tubo flexible (loc-line). El clip de dedo oxímetro de pulso se adjunta a un dedo y se fija con cinta adhesiva. Procedimiento de escaneado Secuencias de pulso: Estructurales: Se utilizan en T1 (imágenes estructurales) que dan una definición clara de la morfología del cerebro de los sujetos. El paciente está en forma pasiva y se mantiene lo más quieto posible durante esta fase, que dura unos 10 minutos. Funcional: Se utilizan secuencias ponderadas en T2 EPI (imágenes funcionales) para mostrar los cambios en la oxigenación de la sangre que se correlacionan con la actividad neuronal. Es durante esta fase que el sujeto está expuesto al jugo, airpuff, y los estímulos auditivos. Conectividad: Se utilizó la difusión de las imágenes ponderadas (DWI) para determinar la conectividad estructural entre las regiones del cerebro. El tema simplemente yace inmóvil durante esta fase.

functional imaging with reinforcement, eyetracking, and physiological monitoring

Usamos imágenes cerebrales funcionales (fMRI) para estudiar los circuitos neuronales que subyacen a la toma de decisiones. Para entender cómo afectan los resultados de los procesos de decisión, las tareas simples de percepción se combinan con un refuerzo apetitivo y aversivo. Sin embargo, el uso de refuerzos, tales como jugos y airpuffs puede crear problemas de resonancia magnética funcional. Entrega de reforzador puede provocar movimiento de la cabeza, lo que crea artefactos en la señal fMRI. Refuerzo también puede conducir a cambios en el ritmo cardíaco y la respiración que son mediadas por vías autonómicas. Los cambios en la frecuencia cardíaca y la respiración puede afectar directamente a la fMRI (BOLD) de la señal en el cerebro y pueden ser confundidos con los cambios de la señal que se deben a la actividad neuronal. En esta presentación se muestran los métodos para la administración de los refuerzos de una manera controlada, para la estabilización de la cabeza, y para medir el pulso y la respiración.

Watch this Scientific Journal Video about Functional Imaging with Reinforcement, Eyetracking, and Physiological Monitoring at JoVE.com

Functional Imaging with Reinforcement, Eyetracking, and Physiological Monitoring

Esta presentación demuestra el uso de resonancia magnética funcional para estudiar los circuitos neuronales que subyacen a la toma de decisiones. Tareas simples de percepción se combinan con los refuerzos del apetito y aversión a investigar cómo los resultados afectan los procesos de decisión.

Imágenes funcionales con Refuerzo, Eyetracking y Monitoreo Fisiológico

We use functional brain imaging (fMRI) to study neural circuits that underlie decision-making.  To understand how outcomes affect decision processes, ...

functional-imaging-with-reinforcement-eyetracking-physiological

Research

JoVE Journal

Biology

11.3K vistas.  Columbia University. Esta presentación demuestra el uso de resonancia magnética funcional para estudiar los circuitos neuronales que subyacen a la toma de decisiones. Tareas simples de percepción se combinan con los refuerzos del apetito y aversión a investigar cómo los resultados afectan los procesos de decisión.

Video: Functional Imaging with Reinforcement, Eyetracking, and Physiological Monitoring

Monitoring Actin Disassembly with Time-lapse Microscopy

Monitoreo de actina desmontaje con la Microscopía de Time-lapse

Investigación

Biología

Labeling hESCs and hMSCs with Iron Oxide Nanoparticles for Non-Invasive in vivo Tracking with MR Imaging

In recent years, stem cell research has led to a better understanding of developmental biology, various diseases and its potential impact on regenerative medicine. A non-invasive method to monitor the transplanted stem cells repeatedly in vivo would greatly enhance our ability to understand the mechanisms that control stem cell death and identify trophic factors and signaling pathways that improve stem cell engraftment. MR imaging has been proven to be an effective tool for the in vivo depiction of stem cells with near microscopic anatomical resolution. In order to detect stem cells with MR, the cells have to be labeled with cell specific MR contrast agents. For this purpose, iron oxide nanoparticles, such as superparamagnetic iron oxide particles (SPIO), are applied, because of their high sensitivity for cell detection and their excellent biocompatibility. SPIO particles are composed of an iron oxide core and a dextran, carboxydextran or starch coat, and function by creating local field inhomogeneities, that cause a decreased signal on T2-weighted MR images. This presentation will demonstrate techniques for labeling of stem cells with clinically applicable MR contrast agents for subsequent non-invasive in vivo tracking of the labeled cells with MR imaging.

For the evaluation of new stem cell therapies it is important to non-invasively track the injected cells in vivo. This video will show you how to label human mesenchymal and embryonic stem cells with iron oxide based contrast agents in vivo for subsequent MR imaging in vivo.

Etiquetado y hESCs hMSCs con nanopartículas de óxido de hierro para no invasivo en el seguimiento in vivo mediante RM

In recent years, stem cell research has led to a better understanding of developmental biology, various diseases and its potential impact on regenerative ...

Labeling Stem Cells with Fluorescent Dyes for non-invasive Detection with Optical Imaging

Optical imaging (OI) is an easy, fast and inexpensive tool for in vivo monitoring of new stem cell based therapies. The technique is based on ex vivo labeling of stem cells with a fluorescent dye, subsequent intravenous injection of the labeled cells and visualization of their accumulation in specific target organs or pathologies. The presented technique demonstrates how we label human mesenchymal stem cells (hMSC) by simple incubation with the lipophilic fluorescent dye DiD (C67H103CIN2O3S) and how we label human embryonic stem cells (hESC) with the FDA approved fluorescent dye Indocyanine Green (ICG). The uptake mechanism is via adherence and diffusion of the lypophilic dye across the phospholipid cell membrane bilayer. The labeling efficiency is usually improved if the cells are incubated with the dye in serum-free media as opposed to incubation in serum-containing media. Furthermore, the addition of the transfection agent Protamine Sulfate significantly improves contrast agent uptake.

This video shows techniques for labeling of human embryonic stem cells and mesenchymal stem cells with fluorescent dyes. This technique can be used for an in vivo tracking of transplanted stem cells with optical imaging and for histopathological correlations with fluorescence microscopy.

Etiquetado de las células madre con tintes fluorescentes para la detección no invasiva con imágenes ópticas

Optical imaging (OI) is an easy, fast and inexpensive tool for in vivo monitoring of new stem cell based therapies. The technique is based on ex vivo ...

Identification of protein complexes with quantitative proteomics in S. cerevisiae

Lipids are the building blocks of cellular membranes that function as barriers and in compartmentalization of cellular processes, and recently, as important intracellular signalling molecules. However, unlike proteins, lipids are small hydrophobic molecules that traffic primarily by poorly described nonvesicular routes, which are hypothesized to occur at membrane contact sites (MCSs). MCSs are regions where the endoplasmic reticulum (ER) makes direct physical contact with a partnering organelle, e.g., plasma membrane (PM). The ER portion of ER-PM MCSs is enriched in lipid-synthesizing enzymes, suggesting that lipid synthesis is directed to these sites and implying that MCSs are important for lipid traffic. Yeast is an ideal model to study ER-PM MCSs because of their abundance, with over 1000 contacts per cell, and their conserved nature in all eukaryotes. Uncovering the proteins that constitute MCSs is critical to understanding how lipids traffic is accomplished in cells, and how they act as signaling molecules. We have found that an ER called Scs2p localize to ER-PM MCSs and is important for their formation. We are focused on uncovering the molecular partners of Scs2p. Identification of protein complexes traditionally relies on first resolving purified protein samples by gel electrophoresis, followed by in-gel digestion of protein bands and analysis of peptides by mass spectrometry. This often limits the study to a small subset of proteins. Also, protein complexes are exposed to denaturing or non-physiological conditions during the procedure. To circumvent these problems, we have implemented a large-scale quantitative proteomics technique to extract unbiased and quantified data. We use stable isotope labeling with amino acids in cell culture (SILAC) to incorporate staple isotope nuclei in proteins in an untagged control strain. Equal volumes of tagged culture and untagged, SILAC-labeled culture are mixed together and lysed by grinding in liquid nitrogen. We then carry out an affinity purification procedure to pull down protein complexes. Finally, we precipitate the protein sample, which is ready for analysis by high-performance liquid chromatography/ tandem mass spectrometry. Most importantly, proteins in the control strain are labeled by the heavy isotope and will produce a mass/ charge shift that can be quantified against the unlabeled proteins in the bait strain. Therefore, contaminants, or unspecific binding can be easily eliminated. By using this approach, we have identified several novel proteins that localize to ER-PM MCSs. Here we present a detailed description of our approach. 

Here we describe a new quantitative proteomics technique for identifying protein complexes in Saccharomyces cerevisiae. In this study, we have used the SILAC method together with affinity purification followed by tandem mass spectrometry to identify with high specificity the binding partners of an ER protein, Scs2p.

Identificación de los complejos de proteínas con la proteómica cuantitativa en S. cerevisiae

Lipids are the building blocks of cellular membranes that function as barriers and in compartmentalization of cellular processes, and recently, as ...

Dissection of Hippocampal Dentate Gyrus from Adult Mouse

The hippocampus is one of the most widely studied areas in the brain because of its important functional role in memory processing and learning, its remarkable neuronal cell plasticity, and its involvement in epilepsy, neurodegenerative diseases, and psychiatric disorders. The hippocampus is composed of distinct regions; the dentate gyrus, which comprises mainly granule neurons, and Ammon's horn, which comprises mainly pyramidal neurons, and the two regions are connected by both anatomic and functional circuits. Many different mRNAs and proteins are selectively expressed in the dentate gyrus, and the dentate gyrus is a site of adult neurogenesis; that is, new neurons are continually generated in the adult dentate gyrus. To investigate mRNA and protein expression specific to the dentate gyrus, laser capture microdissection is often used. This method has some limitations, however, such as the need for special apparatuses and complicated handling procedures. In this video-recorded protocol, we demonstrate a dissection technique for removing the dentate gyrus from adult mouse under a stereomicroscope. Dentate gyrus samples prepared using this technique are suitable for any assay, including transcriptomic, proteomic, and cell biology analyses. We confirmed that the dissected tissue is dentate gyrus by conducting real-time PCR of dentate gyrus-specific genes, tryptophan 2,3-dioxygenase (TDO2) and desmoplakin (Dsp), and Ammon's horn enriched genes, Meis-related gene 1b (Mrg1b) and TYRO3 protein tyrosine kinase 3 (Tyro3). The mRNA expressions of TDO2 and Dsp in the dentate gyrus samples were detected at obviously higher levels, whereas Mrg1b and Tyro3 were lower levels, than those in the Ammon's horn samples. To demonstrate the advantage of this method, we performed DNA microarray analysis using samples of whole hippocampus and dentate gyrus. The mRNA expression of TDO2 and Dsp, which are expressed selectively in the dentate gyrus, in the whole hippocampus of alpha-CaMKII+/- mice, exhibited 0.037 and 0.10-fold changes compared to that of wild-type mice, respectively. In the isolated dentate gyrus, however, these expressions exhibited 0.011 and 0.021-fold changes compared to that of wild-type mice, demonstrating that gene expression changes in dentate gyrus can be detected with greater sensitivity. Taken together, this convenient and accurate dissection technique can be reliably used for studies focused on the dentate gyrus.

A dissection technique for removal of the dentate gyrus from adult mouse under a stereomicroscope was demonstrated in this video-recorded protocol.

La disección de Gyrus dentado del hipocampo de ratones adultos

The hippocampus is one of the most widely studied areas in the brain because of its important functional role in memory processing and learning, its ...

Monitoring Heart Function in Larval Drosophila melanogaster for Physiological Studies

We present various methods to record cardiac function in the larval Drosophila. The approaches allow heart rate to be measured in unrestrained and restrained whole larvae. For direct control of the environment around the heart another approach utilizes the dissected larvae and removal of the internal organs in order to bathe the heart in desired compounds. The exposed heart also allows membrane potentials to be monitored which can give insight of the ionic currents generated by the myocytes and for electrical conduction along the heart tube. These approaches have various advantages and disadvantages for future experiments that are discussed. The larval heart preparation provides an additional model besides the Drosophila skeletal NMJ to investigate the role of intracellular calcium regulation on cellular function. Learning more about the underlying ionic currents that shape the action potentials in myocytes in various species, one can hope to get a handle on the known ionic dysfunctions associated to specific genes responsible for various diseases in mammals.

Monitoring Heart Function in Larval Drosophila melanogaster for Physiological Studies

We present various ways to monitor heart function in the larva of Drosophila for assessing questions dealing with the function of gap junctions, ion channel mutations, modulation of pacemaker activity and pharmacological studies.

Monitorización de la función del corazón en las larvas Drosophila melanogaster Para los estudios fisiológicos

We present various methods to record cardiac function in the larval Drosophila. The approaches allow heart rate to be measured in unrestrained  and  ...

Long-term Lethal Toxicity Test with the Crustacean Artemia franciscana

Our research activities target the use of biological methods for the evaluation of environmental quality, with particular reference to saltwater/brackish water and sediment. The choice of biological indicators must be based on reliable scientific knowledge and, possibly, on the availability of standardized procedures. In this article, we present a standardized protocol that used the marine crustacean Artemia to evaluate the toxicity of chemicals and/or of marine environmental matrices. Scientists propose that the brine shrimp (Artemia) is a suitable candidate for the development of a standard bioassay for worldwide utilization. A number of papers have been published on the toxic effects of various chemicals and toxicants on brine shrimp (Artemia). The major advantage of this crustacean for toxicity studies is the overall availability of the dry cysts; these can be immediately used in testing and difficult cultivation is not demanded1,2. Cyst-based toxicity assays are cheap, continuously available, simple and reliable and are thus an important answer to routine needs of toxicity screening, for industrial monitoring requirements or for regulatory purposes3. The proposed method involves the mortality as an endpoint. The numbers of survivors were counted and percentage of deaths were calculated. Larvae were considered dead if they did not exhibit any internal or external movement during several seconds of observation4. This procedure was standardized testing a reference substance (Sodium Dodecyl Sulfate); some results are reported in this work. This article accompanies a video that describes the performance of procedural toxicity testing, showing all the steps related to the protocol.

Long-term Lethal Toxicity Test with the Crustacean Artemia franciscana

This study concerns the development and standardization of a valuable methodological protocol to determine long-term (14 days) lethal toxicity exerted by chemical substances, industrial wastewater or sewage and liquid environmental samples on the saltwater crustacean, Artemia franciscana.

A largo plazo Prueba de toxicidad letal con el Crustáceo Artemia franciscana</em

Our research activities target the use of biological methods for the evaluation of environmental quality, with particular reference to saltwater/brackish ...

Mapping the After-effects of Theta Burst Stimulation on the Human Auditory Cortex with Functional Imaging

Auditory cortex pertains to the processing of sound, which is at the basis of speech or music-related processing1. However, despite considerable recent progress, the functional properties and lateralization of the human auditory cortex are far from being fully understood. Transcranial Magnetic Stimulation (TMS) is a non-invasive technique that can transiently or lastingly modulate cortical excitability via the application of localized magnetic field pulses, and represents a unique method of exploring plasticity and connectivity. It has only recently begun to be applied to understand auditory cortical function 2. 
An important issue in using TMS is that the physiological consequences of the stimulation are difficult to establish. Although many TMS studies make the implicit assumption that the area targeted by the coil is the area affected, this need not be the case, particularly for complex cognitive functions which depend on interactions across many brain regions 3. One solution to this problem is to combine TMS with functional Magnetic resonance imaging (fMRI). The idea here is that fMRI will provide an index of changes in brain activity associated with TMS. Thus, fMRI would give an independent means of assessing which areas are affected by TMS and how they are modulated 4. In addition, fMRI allows the assessment of functional connectivity, which represents a measure of the temporal coupling between distant regions. It can thus be useful not only to measure the net activity modulation induced by TMS in given locations, but also the degree to which the network properties are affected by TMS, via any observed changes in functional connectivity.
Different approaches exist to combine TMS and functional imaging according to the temporal order of the methods. Functional MRI can be applied before, during, after, or both before and after TMS. Recently, some studies interleaved TMS and fMRI in order to provide online mapping of the functional changes induced by TMS 5-7. However, this online combination has many technical problems, including the static artifacts resulting from the presence of the TMS coil in the scanner room, or the effects of TMS pulses on the process of MR image formation. But more importantly, the loud acoustic noise induced by TMS (increased compared with standard use because of the resonance of the scanner bore) and the increased TMS coil vibrations (caused by the strong mechanical forces due to the static magnetic field of the MR scanner) constitute a crucial problem when studying auditory processing. 
This is one reason why fMRI was carried out before and after TMS in the present study. Similar approaches have been used to target the motor cortex 8,9, premotor cortex 10, primary somatosensory cortex 11,12 and language-related areas 13, but so far no combined TMS-fMRI study has investigated the auditory cortex. The purpose of this article is to provide details concerning the protocol and considerations necessary to successfully combine these two neuroscientific tools to investigate auditory processing. 
Previously we showed that repetitive TMS (rTMS) at high and low frequencies (resp. 10 Hz and 1 Hz) applied over the auditory cortex modulated response time (RT) in a melody discrimination task 2. We also showed that RT modulation was correlated with functional connectivity in the auditory network assessed using fMRI: the higher the functional connectivity between left and right auditory cortices during task performance, the higher the facilitatory effect (i.e. decreased RT) observed with rTMS. However those findings were mainly correlational, as fMRI was performed before rTMS. Here, fMRI was carried out before and immediately after TMS to provide direct measures of the functional organization of the auditory cortex, and more specifically of the plastic reorganization of the auditory neural network occurring after the neural intervention provided by TMS. 
Combined fMRI and TMS applied over the auditory cortex should enable a better understanding of brain mechanisms of auditory processing, providing physiological information about functional effects of TMS. This knowledge could be useful for many cognitive neuroscience applications, as well as for optimizing therapeutic applications of TMS, particularly in auditory-related disorders.

Auditory processing is the basis of speech and music-related processing. Transcranial Magnetic Stimulation (TMS) has been used successfully to study cognitive, sensory and motor systems but has rarely been applied to audition. Here we investigated TMS combined with functional Magnetic Resonance Imaging to understand the functional organization of auditory cortex.

Mapeo de las secuelas de la theta ráfaga de estimulación sobre la corteza auditiva humana con imágenes funcionales

Auditory cortex pertains to the processing of sound, which is at the basis of speech or music-related processing1. However, despite considerable recent ...

Functional Imaging of Brown Fat in Mice with 18F-FDG micro-PET/CT

Brown adipose tissue (BAT) differs from white adipose tissue (WAT) by its discrete location and a brown-red color due to rich vascularization and high density of mitochondria. BAT plays a major role in energy expenditure and non-shivering thermogenesis in newborn mammals as well as the adults 1. BAT-mediated thermogenesis is highly regulated by the sympathetic nervous system, predominantly via &beta; adrenergic receptor 2, 3. Recent studies have shown that BAT activities in human adults are negatively correlated with body mass index (BMI) and other diabetic parameters 4-6. BAT has thus been proposed as a potential target for anti-obesity/anti-diabetes therapy focusing on modulation of energy balance 6-8. While several cold challenge-based positron emission tomography (PET) methods are established for detecting human BAT 9-13, there is essentially no standardized protocol for imaging and quantification of BAT in small animal models such as mice. Here we describe a robust PET/CT imaging method for functional assessment of BAT in mice. Briefly, adult C57BL/6J mice were cold treated under fasting conditions for a duration of 4 hours before they received one dose of 18F-Fluorodeoxyglucose (FDG). The mice were remained in the cold for one additional hour post FDG injection, and then scanned with a small animal-dedicated micro-PET/CT system. The acquired PET images were co-registered with the CT images for anatomical references and analyzed for FDG uptake in the interscapular BAT area to present BAT activity. This standardized cold-treatment and imaging protocol has been validated through testing BAT activities during pharmacological interventions, for example, the suppressed BAT activation by the treatment of &beta;-adrenoceptor antagonist propranolol 14, 15, or the enhanced BAT activation by &beta;3 agonist BRL37344 16. The method described here can be applied to screen for drugs/compounds that modulate BAT activity, or to identify genes/pathways that are involved in BAT development and regulation in various preclinical and basic studies.

Functional Imaging of Brown Fat in Mice with 18F-FDG micro-PET/CT

A method of functional imaging of mouse brown adipose tissue (BAT) is described in which cold-stimulated uptake of 18F-Fluorodeoxyglucose (FDG) in BAT is non-invasively assessed with a standardized micro-PET/CT protocol. This method is robust and sensitive to detect differences in BAT activities in mouse models.

Imagen funcional de grasa marrón en ratones con FDG micro-PET/CT

Brown adipose tissue (BAT) differs from white adipose  tissue (WAT) by its discrete location and a brown-red color due to rich  vascularization and high ...

Concurrent EEG and Functional MRI Recording and Integration Analysis for Dynamic Cortical Activity Imaging

Electroencephalography (EEG) and functional magnetic resonance imaging (fMRI) are two of the fundamental noninvasive methods for identifying brain activity. Multimodal methods have sought to combine the high temporal resolution of EEG with the spatial precision of fMRI, but the complexity of this approach is currently in need of improvement. The protocol presented here describes the recently developed spatiotemporal fMRI-constrained EEG source imaging method, which seeks to rectify source biases and improve EEG-fMRI source localization through the dynamic recruitment of fMRI sub-regions. The process begins with the collection of multimodal data from concurrent EEG and fMRI scans, the generation of 3D cortical models, and independent EEG and fMRI processing. The processed fMRI activation maps are then split into multiple priors, according to their location and surrounding area. These are taken as priors in a two-level hierarchical Bayesian algorithm for EEG source localization. For each window of interest (defined by the operator), specific segments of the fMRI activation map will be identified as active to optimize a parameter known as model evidence. These will be used as soft constraints on the identified cortical activity, increasing the specificity of the multimodal imaging method by reducing cross-talk and avoiding erroneous activity in other conditionally active fMRI regions. The method generates cortical maps of activity and time-courses, which may be taken as final results, or used as a basis for further analyses (analyses of correlation, causation, etc.) While the method is somewhat limited by its modalities (it will not find EEG-invisible sources), it is broadly compatible with most major processing software, and is suitable for most neuroimaging studies.

An EEG-fMRI multimodal imaging method, known as the spatiotemporal fMRI-constrained EEG source imaging method, is described here. The presented method employs conditionally-active fMRI sub-maps, or priors, to guide EEG source localization in a manner that improves spatial specificity and limits erroneous results.