Small-animal System Research Articles

Small-Animal Molecular Imaging for Preclinical Cancer Research: .μPET and μ.SPECT.

Due to different sizes of humans and rodents, the performance of clinical imaging devices is not enough for a scientifically reliable evaluation in mice and rats; therefore dedicated small-animal systems with a much higher sensitivity and spatial resolution, compared to the ones used in humans, are required. Smallanimal imaging represents a cutting-edge research method able to approach an enormous variety of pathologies in which animal models of disease may be used to elucidate the mechanisms underlying the human condition and/or to allow a translational pharmacological (or other) evaluation of therapeutic tools. Molecular imaging, avoiding animal sacrifice, permits repetitive (i.e. longitudinal) studies on the same animal which becomes its own control. In this way also the over time evaluation of disease progression or of the treatment response is enabled. Many different rodent models have been applied to study almost all kind of human pathologies or to experiment a wide series of drugs and/or other therapeutic instruments. In particular, relevant information has been achieved in oncology by in vivo neoplastic phenotypes, obtained through procedures such as subcutaneous tumor grafts, surgical transplantation of solid tumor, orthotopic injection of tumor cells into specific organs/sites of interest, genetic modification of animals to promote tumor-genesis; in this way traditional or innovative treatments, also including gene therapy, of animals with a cancer induced by a known carcinogen may be experimented. Each model has its own disadvantage but, comparing different studies, it is possible to achieve a panoramic and therefore substantially reliable view on the specific subject. Small-animal molecular imaging has become an invaluable component of modern biomedical research that will gain probably an increasingly important role in the next few years.

Design and Verification of Novel Low-Cost MR-Guided Small-Animal Stereotactic System

Stereotactic systems have been used for decades to position surgical instruments in important preclinical and clinical studies. We developed and verified a low-cost magnetic-resonance-guided system for accurate preclinical stereotactic operations on small animals. The system consists of both commercially available and custom components, including in-house software written in MATLAB to register coordinate spaces. The software registers target the image-space coordinates from magnetic resonance imaging to the physical space through a rigid-body transform. The transform uses singular value decomposition to determine rotation and translation matrices that are optimal in a least squares sense. Needle positioning and core biopsy studies were performed to verify the accuracy of the system. The needle positioning study demonstrated a sub-millimeter positioning error of 0.294 ± 0.156 mm while using a 26 s gauge Hamilton needle. Biopsy studies demonstrated >94 % success in accessing sub-millimeter stereotactic targets embedded in agarose gel phantoms. These favorable results confirm that despite its relatively low hardware cost (US$18,730) and easy assembly, the system is able to consistently access stereotactic targets on the spatial scale of a small animal.

Infection with the Persistent Murine Norovirus Strain MNV-S99 Suppresses IFN-Beta Release and Activation of Stat1 In Vitro.

Norovirus infection is the main cause of epidemic non-bacterial gastroenteritis in humans. Although human norovirus (HuNoV) infection is self-limiting, it can persist for extended periods of time in immune deficient patients. Due to the lack of robust cell culture and small animal systems, little is known about HuNoV pathogenicity. However, murine norovirus (MNV) can be propagated in cell culture and is used as a model to study norovirus infection. Several MNV are known to persist in mice. In this study, we show that the MNV strain MNV-S99 persists in wild type inbred (C57BL/6J) mice over a period of at least 5 weeks post infection. Viral RNA was detectable in the jejunum, ileum, cecum, and colon, with the highest titers in the colon and cecum. To characterize the effect of MNV-S99 on the innate immune response, Stat1 phosphorylation and IFN-β production were analyzed and compared to the non-persistent strain MNV-1.CW3. While MNV-S99 and MNV-1.CW3 showed comparable growth characteristics in vitro, Stat1 phosphorylation and IFN-β release is strongly decreased after infection with MNV-S99 compared to MNV-1.CW3. In conclusion, our results show that unlike MNV-1.CW3, MNV-S99 establishes a persistent infection in mice, possibly due to interfering with the innate immune response.

Date (Phoenix dactylifera) Polyphenolics and Other Bioactive Compounds: A Traditional Islamic Remedy's Potential in Prevention of Cell Damage, Cancer Therapeutics and Beyond.

This review analyzes current studies of the therapeutic effects of Phoenix dactylifera, or date palm fruit, on the physiologic system. Specifically, we sought to summarize the effects of its application in preventing cell damage, improving cancer therapeutics and reducing damage caused by conventional chemotherapy. Phoenix dactylifera exhibits potent anti-oxidative properties both in vitro and in vivo. This allows the fruit to prevent depletion of intrinsic protection from oxidative cell damage and assist these defense systems in reducing cell damage. Macroscopically, this mechanism may be relevant to the prevention of various adverse drug events common to chemotherapy including hepatotoxicity, nephrotoxicity, gastrotoxicity, and peripheral neuropathy. While such effects have only been studied in small animal systems, research suggests a potential application to more complex mammalian systems and perhaps a solution to some problems of chemotherapy in hepato-compromised and nephro-compromised patients.

Structural alterations in the rat brain and behavioral impairment after status epilepticus: An MRI study

Temporal lobe epilepsy (TLE) is one of the most common neurologic disorders often associated with behavioral impairments and cognitive deficit. Lithium–pilocarpine model of seizures in rodents reproduces many features of human convulsive status epilepticus (SE) and subsequent TLE. In this study, we have investigated changes in the rat brain after lithium–pilocarpine SE using a high-field MRI system for small animals in early and chronic periods after SE. We have studied the relationship between T2 relaxation time measured in these periods and the development of behavioral exploratory response to novelty and habituation in the open field test. A significant increase in T2 in the hippocampus and associated structures was found 2days after SE and practically resolved by day seven, while an increase in T2 in the parietal and prefrontal cortex appeared 30days after SE. High T2 values in the parietal cortex and thalamus on day two after SE were associated with an increased mortality risk. A substantial variability in T2 relaxation time was observed in the hippocampus and amygdala 30days after SE. Rats survived after SE showed locomotor hyperactivity and disruption of long-term habituation in the open field test carried out 5weeks after the seizures. Interestingly, T2 in the amygdala 30days after SE had a strong correlation with hyperactivity in the novel open field. Therefore, the amygdala damage may be an important factor in the development of hyperactivity in the chronic period after SE.

Meningiomas em cães: aspectos clínicos, histopatológicos e imuno-histoquímicos

Resumo: As neoplasias no sistema nervoso central (SNC) de animais de companhia são frequentemente diagnosticadas, no entanto dados sobre prevalência são escassos. O objetivo deste estudo foi avaliar retrospectivamente a ocorrência de neoplasias primárias de SNC em cães atendidos em um Hospital-Escola Veterinário e descrever aspectos clínicos, histopatológicos e imuno-histoquímicos dos tumores mais frequentes. Quatorze casos (prevalência de 0,27%) de neoplasias primárias de SNC foram identificados no período de 1998 a 2013 e destes, 11 tiveram o diagnóstico de meningiomas. A idade média dos animais com meningioma foi 10 anos, sendo machos (7/11) e a raça Boxer (3/11) os mais afetados. Sete meningiomas eram espinhais e quatro intracranianos, sendo os principais sinais clínicos alteração na locomoção e convulsões, respectivamente. Metástase pulmonar ocorreu em dois casos. Em seis animais com meningioma espinhal foi realizada a mielografia, sendo que em um também foi realizada a tomografia. Em todos os casos os exames foram efetivos na visualização de desvio ou interrupção da coluna de contraste, com alterações sugestivas da presença de massa. Em cinco animais realizou-se cirurgia exploratória visando a confirmação da suspeita clínica ou retirada da massa, sendo que a sobrevida variou de 85 a 960 dias. Na avaliação histopatológica, os meningiomas foram classificados em transicional (4/11), meningotelial (2/11), papilar (2/11), angiomatoso (1/11), microcístico (1/11) e anaplásico (1/11). Destes, oito (8/11) apresentaram marcação positiva para tricrômio de Masson e um para vermelho congo nas técnicas histoquímicas. No painel imuno-histoquímico, todos os casos apresentaram imunomarcação positiva para vimentina, mas imunomarcação negativa para fator VIII e p53. A imunomarcação para S100 (6/11), GFAP (5/11) e pancitoqueratina (3/11) foi de intensidade variável. Na graduação histológica, dez meningiomas eram grau I e um grau III. O índice médio de proliferação celular foi de 3,2 figuras de mitose e 3,4% avaliando a expressão de Ki-67. Os resultados confirmaram que os meningiomas são a neoplasia primária de SNC mais prevalente em cães, com variação nos subtipos histológicos. A caracterização histoquímica e imuno-histoquímica contribuiu com a determinação do diagnóstico, no entanto, estudos envolvendo a expressão de outros genes são necessários para auxiliar no prognóstico dessas neoplasias.

Performance characterization of a new CZT-based preclinical SPECT system: a comparative study of different collimators

Triumph X-SPECT is a newly released CZT-based preclinical small-animal SPECT system with interchangeable collimators. The purpose of this work was to evaluate and systematically compare the imaging performances of three different collimators in the CZT-based preclinical small-animal system: a single-pinhole collimator (SPH), a multi-pinhole collimator (MPH) and a parallel-hole collimator. We measured the spatial resolutions and sensitivities of the three collimators with 99mTc sources, considering three distinct energy window widths (5, 10, and 20%), and used the NEMA NU4-2008 Image Quality phantom to test the imaging performance of the three collimators in terms of uniformity and spill-over ratio (SOR) for each energy window. With a 10% energy window width at a radius of rotation (ROR) of 30 mm, the system resolution of the SPH, MPH and parallel-hole collimators was 0.715, 0.855 and 3.270 mm FWHM, respectively. For the same energy window, the sensitivity of the system with SPH, MPH and parallel-hole collimators was 32.860, 152.514 and 49.205 counts/sec/MBq at a 100 mm source-to-detector distance and 6.790, 33.376 and 49.038 counts/sec/MBq at a 130 mm source-to-detector distance, respectively. The image noise and SORair for the three collimators were 20.137, 12.278 and 11.232 (%STDunif) and 0.106, 0.140 and 0.161, respectively. Overall, the results show that the SPH had better spatial resolution than the other collimators. The MPH had the highest sensitivity at 100 mm source-to-collimator distance, and the parallel-hole collimator had the highest sensitivity at 130 mm-source-to-detector distance. Therefore, the proper collimator for Triumph X-SPECT system must be determined by the task. These results provide valuable reference data and insight into the imaging performance of various collimators in CZT-based preclinical small-animal SPECT.

Performance characterization of a new CZT-based preclinical SPECT system: a comparative study of different collimators

Triumph X-SPECT is a newly released CZT-based preclinical small-animal SPECT system with interchangeable collimators. The purpose of this work was to evaluate and systematically compare the imaging performances of three different collimators in the CZT-based preclinical small-animal system: a single-pinhole collimator (SPH), a multi-pinhole collimator (MPH) and a parallel-hole collimator. We measured the spatial resolutions and sensitivities of the three collimators with 99mTc sources, considering three distinct energy window widths (5, 10, and 20%), and used the NEMA NU4-2008 Image Quality phantom to test the imaging performance of the three collimators in terms of uniformity and spill-over ratio (SOR) for each energy window. With a 10% energy window width at a radius of rotation (ROR) of 30 mm, the system resolution of the SPH, MPH and parallel-hole collimators was 0.715, 0.855 and 3.270 mm FWHM, respectively. For the same energy window, the sensitivity of the system with SPH, MPH and parallel-hole collimators was 32.860, 152.514 and 49.205 counts/sec/MBq at a 100 mm source-to-detector distance and 6.790, 33.376 and 49.038 counts/sec/MBq at a 130 mm source-to-detector distance, respectively. The image noise and SORair for the three collimators were 20.137, 12.278 and 11.232 (%STDunif) and 0.106, 0.140 and 0.161, respectively. Overall, the results show that the SPH had better spatial resolution than the other collimators. The MPH had the highest sensitivity at 100 mm source-to-collimator distance, and the parallel-hole collimator had the highest sensitivity at 130 mm-source-to-detector distance. Therefore, the proper collimator for Triumph X-SPECT system must be determined by the task. These results provide valuable reference data and insight into the imaging performance of various collimators in CZT-based preclinical small-animal SPECT.

Changes in inflammatory response during and after cardiopulmonary bypass using a rat extracorporeal circulation model.

Cardiopulmonary bypass (CPB) is indispensable for cardiac surgery. Since the ethical and technical difficulties associated with clinical research limit the acquisition of new knowledge, it is desirable to have a miniature extracorporeal circulation (ECC) system for small animals. We aimed to establish a miniature ECC system and apply the system to the rat for investigating systemic inflammatory response changes during and after CPB. The ECC system consisted of a membranous oxygenator (polypropylene, 0.03 m(2)), tubing line (polyvinyl chloride) and a roller pump. Priming volume of this system is only 7 ml. Rats were divided into the SHAM (received surgical preparation only without CPB) group and the CPB group. ECC pump flow was initiated and maintained at 70 ml/kg/min. We measured the serum cytokine levels of tumor necrosis factor-α, interleukin (IL)-6, and IL-10, and biochemical markers (lactate dehydrogenase, aspartate aminotransferase and alanine aminotransferase) before, 30, and 60 min after the initiation of CPB, in addition, 30, 60, and 120 min after the CPB weaning. During CPB, blood pressure and hemoglobin were maintained around 80 mmHg and 10g/dl, the serum cytokine levels and biochemical markers were significantly elevated in the CPB group compared with the SHAM group. These data suggest that CPB promotes organ damage and a systemic inflammatory response. This rat ECC model is considered to be equivalent to the already established human CPB and useful for studying the mechanism of pathophysiological changes during and after CPB.

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 µm, 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 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 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.

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 µm, 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 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 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.

Multi-Wavelength Photoacoustic Visualization of High Intensity Focused Ultrasound Lesions.

High intensity focused ultrasound (HIFU) thermal therapies are limited by deficiencies in existing image-guidance techniques. Previous studies using single-wavelength photoacoustic (PA) imaging have demonstrated that HIFU lesions generate contrast with respect to native tissues but have not sufficiently assessed lesion extent. The purpose of this study is to demonstrate feasibility of characterization of in vitro HIFU ablation lesion dimensions using 3D multi-wavelength PA imaging. Fresh porcine cardiac and liver tissue samples were embedded in agar phantoms and ablated using a 2.5 MHz small-animal HIFU system. Both 2D and 3D multi-wavelength photoacoustic-ultrasonic (PAUS) scans were performed in the near-infrared (NIR) range to characterize the change in the absorption spectrum of tissues following ablation and were compared to stained gross pathology to assess treatment margins and lesion extent. Comprehensive 2D multi-wavelength PA imaging yielded a spectrum in ablated tissue that did not display the characteristic local maximum in the optical absorption spectrum of deoxy-hemoglobin (Hb) near 760 nm. Two-dimensional tissue characterization map (TCM) images reconstructed from 3D TCM volumes reliably characterized lesion area and showed >70% area agreement with stained gross pathology. In addition, tissue samples were heated via water bath and concurrently interrogated with 2D PAUS imaging. PA signal exhibited an initial amplitude increase across all wavelengths, corresponding to an initial temperature increase, before then exhibiting a spectral change. This study suggests that multi-wavelength PA imaging has potential to obtain accurate characterization of HIFU lesion extent and may be better suited to guide HIFU ablation therapies during clinical treatments than single-wavelength methods.

Design optimization of a small-animal SPECT system using LGSO continuous crystals and micro parallel-hole collimators

A small-animal single photon emission computed tomography (SPECT) system having compact size and low cost was designed using monolithic LGSO scintillation crystals and micro parallel-hole collimators through Monte Carlo simulations. A spatial resolution of ~1 mm and a sensitivity of ~100 cps/MBq were achieved with a four-head SPECT system. A hot rod with a diameter of 1.0 mm was resolved in the SPECT image of the ultra-micro hot spot phantom. Using a thin monolithic crystal and micro collimator, we achieved high spatial resolution and high sensitivity.

Adaptation, Commissioning, and Evaluation of a 3D Treatment Planning System for High-Resolution Small-Animal Irradiation.

Although spatially precise systems are now available for small-animal irradiations, there are currently limited software tools available for treatment planning for such irradiations. We report on the adaptation, commissioning, and evaluation of a 3-dimensional treatment planning system for use with a small-animal irradiation system. The 225-kV X-ray beam of the X-RAD 225Cx microirradiator (Precision X-Ray) was commissioned using both ion-chamber and radiochromic film for 10 different collimators ranging in field size from 1 mm in diameter to 40 × 40 mm(2) A clinical 3-dimensional treatment planning system (Metropolis) developed at our institution was adapted to small-animal irradiation by making it compatible with the dimensions of mice and rats, modeling the microirradiator beam orientations and collimators, and incorporating the measured beam data for dose calculation. Dose calculations in Metropolis were verified by comparison with measurements in phantoms. Treatment plans for irradiation of a tumor-bearing mouse were generated with both the Metropolis and the vendor-supplied software. The calculated beam-on times and the plan evaluation tools were compared. The dose rate at the central axis ranges from 74 to 365 cGy/min depending on the collimator size. Doses calculated with Metropolis agreed with phantom measurements within 3% for all collimators. The beam-on times calculated by Metropolis and the vendor-supplied software agreed within 1% at the isocenter. The modified 3-dimensional treatment planning system provides better visualization of the relationship between the X-ray beams and the small-animal anatomy as well as more complete dosimetric information on target tissues and organs at risk. It thereby enhances the potential of image-guided microirradiator systems for evaluation of dose-response relationships and for preclinical experimentation generally.

Experimental system to detect a labeled cell monolayer in a microfluidic environment.

To investigate the feasibility of detecting a living cell monolayer labeled with gadoterate (Gd-DOTA) in a microfluidic environment, by micromagnetic resonance imaging (MRI) in a 2.35T small-animal system. The development of new targeted contrast agents (CAs) requires proof-of-concept studies in order to establish the detectability of the CA and to predict the role of biodistribution in its uptake mechanisms. A promising approach is to carefully mimic the in vivo pharmacokinetic context with reduced experimental complexity compared to in vivo situations. A dedicated experimental system was built by combining a microfluidic slide and a radiofrequency probe based on a 6 mm diameter multiturn transmission-line resonator. Adherent KB cells were incubated with different concentrations of Gd. MRI data were acquired at 2.35T with a 3D gradient echo and a resolution of 12.4 μm perpendicular to the cell layer. The longitudinal relaxation rate, R1 , was measured as a function of the amount of Gd internalized by the cells. R1 measurements for different Gd concentrations per cell were performed using data with an signal-to-noise ratio (SNR) of 100. Relaxation-rate variations ΔR1 of 0.035 s(-1) were measured. A quenching effect was observed at Gd concentrations above 20 fmol/cell. Our results suggest that this dedicated experimental system is suitable for specifically assessing new high-relaxivity targeted CAs under real-time uptake conditions.

Validation of nanobody and antibody based in vivo tumor xenograft NIRF-imaging experiments in mice using ex vivo flow cytometry and microscopy.

This protocol outlines the steps required to perform ex vivo validation of in vivo near-infrared fluorescence (NIRF) xenograft imaging experiments in mice using fluorophore labelled nanobodies and conventional antibodies.First we describe how to generate subcutaneous tumors in mice, using antigen-negative cell lines as negative controls and antigen-positive cells as positive controls in the same mice for intraindividual comparison. We outline how to administer intravenously near-infrared fluorophore labelled (AlexaFluor680) antigen-specific nanobodies and conventional antibodies. In vivo imaging was performed with a small-animal NIRF-Imaging system. After the in vivo imaging experiments the mice were sacrificed. We then describe how to prepare the tumors for parallel ex vivo analyses by flow cytometry and fluorescence microscopy to validate in vivo imaging results.The use of the near-infrared fluorophore labelled nanobodies allows for non-invasive same day imaging in vivo. Our protocols describe the ex vivo quantification of the specific labeling efficiency of tumor cells by flow cytometry and analysis of the distribution of the antibody constructs within the tumors by fluorescence microscopy. Using near-infrared fluorophore labelled probes allows for non-invasive, economical in vivo imaging with the unique ability to exploit the same probe without further secondary labelling for ex vivo validation experiments using flow cytometry and fluorescence microscopy.

Validation of Nanobody and Antibody Based <em>In Vivo</em> Tumor Xenograft NIRF-imaging Experiments in Mice Using <em>Ex Vivo</em> Flow Cytometry and Microscopy

This protocol outlines the steps required to perform ex vivo validation of in vivo near-infrared fluorescence (NIRF) xenograft imaging experiments in mice using fluorophore labelled nanobodies and conventional antibodies. First we describe how to generate subcutaneous tumors in mice, using antigen-negative cell lines as negative controls and antigen-positive cells as positive controls in the same mice for intraindividual comparison. We outline how to administer intravenously near-infrared fluorophore labelled (AlexaFluor680) antigen-specific nanobodies and conventional antibodies. In vivo imaging was performed with a small-animal NIRF-Imaging system. After the in vivo imaging experiments the mice were sacrificed. We then describe how to prepare the tumors for parallel ex vivo analyses by flow cytometry and fluorescence microscopy to validate in vivo imaging results. The use of the near-infrared fluorophore labelled nanobodies allows for non-invasive same day imaging in vivo. Our protocols describe the ex vivo quantification of the specific labeling efficiency of tumor cells by flow cytometry and analysis of the distribution of the antibody constructs within the tumors by fluorescence microscopy. Using near-infrared fluorophore labelled probes allows for non-invasive, economical in vivo imaging with the unique ability to exploit the same probe without further secondary labelling for ex vivo validation experiments using flow cytometry and fluorescence microscopy.

Non-invasive parenchymal, vascular and metabolic high-frequency ultrasound and photoacoustic rat deep brain imaging.

Photoacoustics and high frequency ultrasound stands out as powerful tools for neurobiological applications enabling high-resolution imaging on the central nervous system of small animals. However, transdermal and transcranial neuroimaging is frequently affected by low sensitivity, image aberrations and loss of space resolution, requiring scalp or even skull removal before imaging. To overcome this challenge, a new protocol is presented to gain significant insights in brain hemodynamics by photoacoustic and high-frequency ultrasounds imaging with the animal skin and skull intact. The procedure relies on the passage of ultrasound (US) waves and laser directly through the fissures that are naturally present on the animal cranium. By juxtaposing the imaging transducer device exactly in correspondence to these selected areas where the skull has a reduced thickness or is totally absent, one can acquire high quality deep images and explore internal brain regions that are usually difficult to anatomically or functionally describe without an invasive approach. By applying this experimental procedure, significant data can be collected in both sonic and optoacoustic modalities, enabling to image the parenchymal and the vascular anatomy far below the head surface. Deep brain features such as parenchymal convolutions and fissures separating the lobes were clearly visible. Moreover, the configuration of large and small blood vessels was imaged at several millimeters of depth, and precise information were collected about blood fluxes, vascular stream velocities and the hemoglobin chemical state. This repertoire of data could be crucial in several research contests, ranging from brain vascular disease studies to experimental techniques involving the systemic administration of exogenous chemicals or other objects endowed with imaging contrast enhancement properties. In conclusion, thanks to the presented protocol, the US and PA techniques become an attractive noninvasive performance-competitive means for cortical and internal brain imaging, retaining a significant potential in many neurologic fields.

Non-invasive Parenchymal, Vascular and Metabolic High-frequency Ultrasound and Photoacoustic Rat Deep Brain Imaging

Photoacoustics and high frequency ultrasound stands out as powerful tools for neurobiological applications enabling high-resolution imaging on the central nervous system of small animals. However, transdermal and transcranial neuroimaging is frequently affected by low sensitivity, image aberrations and loss of space resolution, requiring scalp or even skull removal before imaging. To overcome this challenge, a new protocol is presented to gain significant insights in brain hemodynamics by photoacoustic and high-frequency ultrasounds imaging with the animal skin and skull intact. The procedure relies on the passage of ultrasound (US) waves and laser directly through the fissures that are naturally present on the animal cranium. By juxtaposing the imaging transducer device exactly in correspondence to these selected areas where the skull has a reduced thickness or is totally absent, one can acquire high quality deep images and explore internal brain regions that are usually difficult to anatomically or functionally describe without an invasive approach. By applying this experimental procedure, significant data can be collected in both sonic and optoacoustic modalities, enabling to image the parenchymal and the vascular anatomy far below the head surface. Deep brain features such as parenchymal convolutions and fissures separating the lobes were clearly visible. Moreover, the configuration of large and small blood vessels was imaged at several millimeters of depth, and precise information were collected about blood fluxes, vascular stream velocities and the hemoglobin chemical state. This repertoire of data could be crucial in several research contests, ranging from brain vascular disease studies to experimental techniques involving the systemic administration of exogenous chemicals or other objects endowed with imaging contrast enhancement properties. In conclusion, thanks to the presented protocol, the US and PA techniques become an attractive noninvasive performance-competitive means for cortical and internal brain imaging, retaining a significant potential in many neurologic fields.

Ressonância magnética no diagnóstico de doença do disco intervertebral em cães – revisão

Magnetic resonance imaging has been significantly used in USA, Canada and Europe for evaluation of diseases affecting the nervous system in small animals. As far as the spine is concerned, the technique can be employed to assess the integrity of intervertebral discs and of structures adjacent to the vertebral body and to the spinal cord. To interpret the images, the radiologist needs to know how to execute the main pulse sequences and image planes. A good technical quality and anatomical knowledge are essential to avoid diagnostic failures and mistakes. The aim of this review is to describe the technical principles, specific pulse sequences and imagen planes when examining the dog spine, as well as the spine anatomy seen through magnetic resonance and the most common findings in the different types of intervertebral disc disease in dogs.