RESUMEN
Miniaturized fluorescence microscopes (miniscopes) have been instrumental to monitor neural signals during unrestrained behavior and their open-source versions have made them affordable. Often, the footprint and weight of open-source miniscopes is sacrificed for added functionality. Here, we present NINscope: a light-weight miniscope with a small footprint that integrates a high-sensitivity image sensor, an inertial measurement unit and an LED driver for an external optogenetic probe. We use it to perform the first concurrent cellular resolution recordings from cerebellum and cerebral cortex in unrestrained mice, demonstrate its optogenetic stimulation capabilities to examine cerebello-cerebral or cortico-striatal connectivity, and replicate findings of action encoding in dorsal striatum. In combination with cross-platform acquisition and control software, our miniscope is a versatile addition to the expanding tool chest of open-source miniscopes that will increase access to multi-region circuit investigations during unrestrained behavior.
Asunto(s)
Microscopía Fluorescente/instrumentación , Red Nerviosa/anatomía & histología , Animales , Conducta Animal , Encéfalo/anatomía & histología , Encéfalo/diagnóstico por imagen , Femenino , Imagenología Tridimensional , Masculino , Ratones Endogámicos C57BL , OptogenéticaRESUMEN
PURPOSE: To investigate continuous distraction osteogenesis (DO) of the nasal bones in a rabbit model, and to compare data from this continuous DO study with data from a previously conducted discontinuous DO study. In addition, radiographic and ultrasonographic bone-fill scores were determined to investigate whether these scores provided reliable predictive value for the amount of new bone formation in the distraction area. MATERIALS AND METHODS: Skeletally mature female New Zealand White rabbits were subjected to distraction of the nasal bones. A custom-made continuous distractor was used to perform automatic non-stop distraction. Bone data were obtained from radiography, ultrasonography, and microcomputed tomography. Data from this experiment were compared with data from a previous study on discontinuous distraction rhythms. RESULTS: Ultrasonographic bone-fill scores correlated significantly to actual bone volume in contrast to radiographic bone-fill scores. Bone volume was significantly higher in the continuous DO group compared with the discontinuous DO groups. CONCLUSION: Continuous distraction resulted in accelerated osteogenesis compared with discontinuous distraction. Furthermore, bone-fill scores based on ultrasonography showed a significant correlation with actual bone volumes.
Asunto(s)
Regeneración Ósea/fisiología , Osteogénesis por Distracción/métodos , Animales , Densidad Ósea/fisiología , Fijadores Externos , Femenino , Modelos Animales , Hueso Nasal/diagnóstico por imagen , Hueso Nasal/patología , Hueso Nasal/cirugía , Osteogénesis/fisiología , Osteogénesis por Distracción/instrumentación , Conejos , Ultrasonografía , Microtomografía por Rayos XRESUMEN
PURPOSE: During retinal pigment epithelium (RPE) and choroid graft translocation in the treatment of patients with exudative age-related macular degeneration, the adhesion of the graft to the translocation instrument complicated its submacular release. Vibration of the instrument improved the release of the graft. This study was conducted to validate the effectiveness of the principle of vibration and to determine the threshold amplitude and frequency required for development of an optimized instrument. METHODS: An experimental in vitro model with fresh porcine RPE-choroid grafts was used. Release of the graft was studied by a masked observer for amplitudes in the range of 0.05 to 1.2 mm and frequencies in the range of 25 to 200 Hz in the horizontal plane. RESULTS: The minimum threshold amplitude required to release the graft was approximately 0.15 mm from a frequency of 100 Hz and higher. CONCLUSIONS: This study confirmed the clinical experience that vibration of an instrument induces the release of the RPE-choroid graft. The minimum threshold amplitude and frequency needed for optimum tissue release were estimated.
Asunto(s)
Coroides/trasplante , Procedimientos Quirúrgicos Oftalmológicos/instrumentación , Epitelio Pigmentado Ocular/trasplante , Vibración , Animales , Modelos Teóricos , Umbral Sensorial , PorcinosRESUMEN
The focal location of atherosclerosis in the vascular tree is correlated with local variations in shear stress. We developed a method to induce defined variations in shear stress in a straight vessel segment of a mouse. To this end, a cylinder with a tapered lumen was placed around the carotid artery, inducing a high shear stress field. Concomitantly, regions of low shear stress and oscillatory shear stress were created upstream and down-stream of the device, respectively. This device was used in mice transgenic for an eNOS3GFP fusion gene. We observed a strong induction of endothelial nitric oxide synthase-green fluorescent protein (eNOS-GFP) mRNA expression in the high shear stress region compared with the other regions (P < .05). Quantification of eNOS-GFP fluorescence or of immunoreactivity to the Golgi complex or to platelet endothelial cell adhesion molecule 1 (PECAM-1) showed an increase in the high shear stress region (P < .05) compared with nontreated carotid arteries. Colocalization of eNOS-GFP with either the Golgi complex or PECAM-1 also responded to alterations of shear stress. In conclusion, we showed a direct response of mRNA and protein expression in vivo to induced variations of shear stress. This model provides the opportunity to study the relationship between shear stress alterations, gene expression, and atherosclerosis.