<ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> تخدير الفئران مع isoflurane (4 ٪ لتحريض ، 1،5-2 ٪ للجراحة) باستخدام IACUC الإجراءات المعتمدة. من المهم أن تستخدم الذيل و / أو يقرص اصبع القدم من أجل ضمان مخدرا تماما الحيوانية. </li><li style=";text-align:right;direction:rtl"> باستخدام الانتهازي القوارض ، وحلق الشعر من الجزء الخلفي من الرقبة حتى العينين. </li><li style=";text-align:right;direction:rtl"> ضع الماوس في إطار التجسيمي ، على مدى بطانية جراحة إعادة تدوير المياه. آمن بقوة الرأس بقضبان الأذن. </li><li style=";text-align:right;direction:rtl"> تطبيق مرهم العين ، من أجل منع العين الحيوان من الجفاف. </li><li style=";text-align:right;direction:rtl"> إدارة ، تحت الجلد ، ديكساميتازون (0.2 ملغ / كلغ) ، وكاربروفين (5 ملغ / كلغ) لمنع تورم في الدماغ و / أو الاستجابة الالتهابية ، على التوالي. </li><li style=";text-align:right;direction:rtl"> قبل البدء في الجراحة ، وتعقيم المنطقة التي تعمل بواسطة المسح الجلد مع ثلاثة الضربات الشديدة بالتناوب من الكحول 70 ٪ وbetadine. </li><li style=";text-align:right;direction:rtl"> تم الانتهاء من جميع الأدوات الجراحية قبل تعقيمها باستخدام حبة تعقيم الزجاج. باستخدام مقص التي تم تعقيمها مع الإيثانول ، إزالة الجلد فوق الجزء العلوي من الجمجمة ، بدءا من قطع أفقي على طول قاعدة الرأس ، تليها اثنين من التخفيضات في الاتجاه منقاري ، ليصل إلى ما يقرب من الجفون ، ثم اثنين من التخفيضات المائل التي تلتقي في خط الوسط. </li><li style=";text-align:right;direction:rtl"> يتم تطبيق قطرة من محلول يدوكائين الإيبينيفرين + في هذه المرحلة على السمحاق لتجنب النزيف أو الألم. مع مشرط ، سحب السمحاق على حواف الجمجمة. أيضا ، يتراجع قليلا في الجهاز العضلي من الجزء الخلفي من الرقبة. </li><li style=";text-align:right;direction:rtl"> كشط بلطف على المنطقة بأكملها معرضة للجمجمة مع مشرط لخلق سطح جاف. هذا مهم جدا ، لأنها سوف تسمح للالغراء التمسك أفضل عند تطبيقها في وقت لاحق. </li><li style=";text-align:right;direction:rtl"> مرة واحدة وقد تم اختياره في موقع التصوير ، واحد هو على استعداد لإنشاء إطار الجمجمة. أولا ، بلطف &quot;لرسم&quot; حلقة من حوالي 4 مم وقطرها مع التدريبات الهوائية الأسنان. </li><li style=";text-align:right;direction:rtl"> بعد حفر طفيفة ، وتطبيق حل الإيبينيفرين يدوكائين + مرة أخرى على سطح الجمجمة. وقف الحفر عند ترك طبقة رقيقة جدا من العظام. بالضغط بلطف على مركز حج القحف أن يشعر كيف أنها تعطي الطريقة ، يمكن للمرء أن يعرف عادة أن يتم التوصل إلى هذه المرحلة. </li><li style=";text-align:right;direction:rtl"> تحت قطرة من المياه المالحة والاستفادة من اتخاذ ترابيق العظم -- هيكل الاسفنجية للعظم -- رفع حج القحف بعيدا من الجمجمة مع ملقط تلميح رقيقة جدا. والمالحة هو المهم ، لأنها سوف تساعد في رفع تصل الجمجمة ونزيف في منع الجافية. </li><li style=";text-align:right;direction:rtl"> تطبيق جلفوم التي تم نقعها في المياه المالحة في السابق لجافية لإيقاف أي نزيف يحدث أحيانا أن صغيرة عند إزالة الجمجمة. </li><li style=";text-align:right;direction:rtl"> بعد تجفيف سطح جافية ، وضمان عدم وجود نزيف ، تكمن برفق زجاج معقمة 5 مم زلة تغطية على رأس الأم الجافية. (ملاحظة : المجموعات الأخرى أيضا وضع قطرة من انخفاض درجة انصهار agarose (1.2 ٪) خلال الجافية ووضع على رأس ساترة للآجار). </li><li style=";text-align:right;direction:rtl"> تطبيق قطرة cyanocrylate مقرها الغراء لنصف الكرة المعاكس على الجمجمة. مع مساعدة من إبرة ، وتطبيق برفق الغراء في جميع أنحاء نافذة مع الحرص على عدم وضعه تحت الزجاج. ويمكن الآن الغراء تطبق في طبقة رقيقة فوق سطح الأرض كله من الجمجمة. </li><li style=";text-align:right;direction:rtl"> بمجرد أن يجف الغراء ، مزيج من الاكريليك الأسنان وتطبيقه في جميع أنحاء سطح الجمجمة ، وتغطي أيضا حافة صغيرة من زلة تغطية ، لضمان الحصول عليها. </li><li style=";text-align:right;direction:rtl"> بعد الحصول على تغطية زلة ، وجعل بئر صغيرة حول الإطار مع الاكريليك الأسنان. أيضا ، تضمين شريط التيتانيوم في الاكريليك الأسنان. سيتم لاحقا هذا الشريط أن تستخدم لتوصيل الماوس بأمان إلى مرحلة من مراحل المجهر للتصوير. فمن المهم لضمان أن شريط غير مستوى ، بحيث يسير بشكل متواز مع النافذة في الجمجمة. ويمكن وضع قطعة من الورق تحت شريط يسمح للشريط على البقاء في حين أن مستوى الاكريليك يصلب. </li><li style=";text-align:right;direction:rtl"> ولا يسمح للالاكريليك لعلاج الأسنان (تصلب) لمدة 10 دقيقة ، وهو الوقت الذي يتم إصلاح شريط التيتانيوم في المكان. وضع الحيوانات في قفص الدافئة حتى يتعافى. </li><li style=";text-align:right;direction:rtl"> بعد التعافي من التخدير ، ويمكن تصوير الحيوان في نفس اليوم. </li></ol>

<ol>
	<li>Svoboda, K., Denk, W., Kleinfeld, D., Tank, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=+In+vivo+dendritic+calcium+dynamics+in+neocortical+pyramidal+neurons."> In vivo dendritic calcium dynamics in neocortical pyramidal neurons.</a> Nature. 85, 161-165 (1997).</li><li>Lendvai, B., Stern, E. A., Chen, B., Svoboda, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experience-dependent+plasticity+of+dendritic+spines+in+the+developing+rat+barrel+cortex+in+vivo.">Experience-dependent plasticity of dendritic spines in the developing rat barrel cortex in vivo.</a> Nature. 404, 876-881 (2000).</li><li>Trachtenberg, J. T., Chen, B., Knott, G. W., Feng, G., Sanes, J. R., Welker, E., Svoboda, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+in+vivo+imaging+of+experience-dependent+synaptic+plasticity+in+adult+cortex.">Long-term in vivo imaging of experience-dependent synaptic plasticity in adult cortex.</a> Nature. 420, 788-794 (2002).</li><li>Portera-Cailliau, C., Weimer, R. M., DePaola, V., Caroni, P., Svoboda, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diverse+modes+of+axon+elaboration+in+the+developing+neocortex.">Diverse modes of axon elaboration in the developing neocortex.</a> PLoS Biol. 3, (2005).</li><li>Holtmaat, A., Wilbrecht, L., Knott, G. W., Welker, E., Svoboda, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experience-dependent+and+cell-type-specific+spine+growth+in+the+neocortex.">Experience-dependent and cell-type-specific spine growth in the neocortex.</a> Nature. 441, 979-983 (2006).</li></ol>

كما بينا في شريط الفيديو والتكميلية في الأرقام ، وإعداد إطار الجمجمة ، جنبا إلى جنب مع استخدام اثنين من الفوتون المجهري ، هو أداة قوية جدا للدراسة في بنية الجسم الحي وظيفة من وظائف القشرة المخية الحديثة. تقنية يتطلب تدريب صارم لتصبح مألوفة مع التشريح ذات الصلة والإجراءات الجراح...

قضبان من التيتانيوم ومصنوعة خصيصا

a craniotomy surgery procedure for chronic brain imaging

تقنيات التصوير أصبحت ذات أهمية متزايدة في دراسة وظائف المخ. من بينها ، برز ثنائي الفوتون المجهري المسح بواسطة أشعة الليزر كطريقة مفيدة للغاية ، لأنه يسمح للدراسة الدماغ سليمة حية. مع الاستعدادات المناسبة ، وهذا الأسلوب يسمح للمراقبة المنطقة القشرية نفس مزمنة ، من دقائق إلى أشهر. في هذا الفيديو ، نظهر استعدادا لتصوير المزمن في الجسم الحي من الدماغ باستخدام اثنين من الفوتون المجهري. كان في البداية هذه التقنية التي بدأها الدكتور كاريل سفوبودا ، وهو الآن في معهد هوارد هيوز الطبي محقق في مزرعة Janelia. ويمكن استخدام مستحضرات مثل واحد هو موضح هنا لتصوير هيكل القشرة المخية الحديثة (على سبيل المثال ، وديناميات الشجيرية ومحواري) ، لتسجيل نشاط الخلايا العصبية باستخدام الكالسيوم الحساسة الأصباغ ، لالقشرية صورة ديناميكية تدفق الدم ، أو للدراسات الذاتية التصوير الضوئي. التصوير العميق للاللحاء الجديد هو ممكن مع أفضل إطار عملية جراحية في الجمجمة. تعمل في ظل ظروف بالغة العقيمة من الممكن تجنب الإصابة بالأمراض ، جنبا إلى جنب مع استخدام العناية القصوى للا تضر الأم الجافية أثناء الجراحة ، سوف يؤدي إلى النوافذ الزجاجية المغطاة ناجحة وطويلة الأمد في الجمجمة.

Watch this Scientific Journal Video about A Craniotomy Surgery Procedure for Chronic Brain Imaging at JoVE.com

A Craniotomy Surgery Procedure for Chronic Brain Imaging

هذا الفيديو وبروتوكول لشرح كيفية زرع زجاج النافذة المغطاة الجمجمة لدى القوارض. ويمكن استخدام هذه المستحضرات لمزمنة في الجسم الحي two الفوتون التصوير من القشرة المخية الحديثة على جداول زمنية لأشهر. ويمكن أيضا استخدامه لأنواع أخرى من التصوير ، والتصوير الضوئي بما في ذلك إشارة جوهرية.

إجراء جراحة حج القحف لتصوير الدماغ المزمن

Imaging techniques are becoming increasingly important in the study brain function. Among them, two-photon laser scanning microscopy has emerged as an ...

a-craniotomy-surgery-procedure-for-chronic-brain-imaging

Research

JoVE Journal

Biology

63.6K Views.  University of California, Los Angeles. هذا الفيديو وبروتوكول لشرح كيفية زرع زجاج النافذة المغطاة الجمجمة لدى القوارض. ويمكن استخدام هذه المستحضرات لمزمنة في الجسم الحي two الفوتون التصوير من القشرة المخية الحديثة على جداول زمنية لأشهر. ويمكن أيضا استخدامه لأنواع أخرى من التصوير ، والتصوير الضوئي بما...

Video: A Craniotomy Surgery Procedure for Chronic Brain Imaging

Application of Light-cured Dental Adhesive Resin for Mounting Electrodes or Microdialysis Probes in Chronic Experiments

In chronic recording experiments, self-curing dental acrylic resins have been used as a mounting base of electrodes or microdialysis-probes. Since these acrylics do not bond to the bone, screws have been used as anchors. However, in small experimental animals like finches or mouse, their craniums are very fragile and can not successfully hold the anchors. In this report, we propose a new application of light-curing dental resins for mounting base of electrodes or microdialysis probes in chronic experiments. This material allows direct bonding to the cranium. Therefore, anchor screws are not required and surgical field can be reduced considerably. Past experiences show that the bonding effect maintains more than 2 months. Conventional resin's window of time when the materials are pliable and workable is a few minutes. However, the window of working time for these dental adhesives is significantly wider and adjustable.

In this report, we propose a new application of light-curing dental resins for mounting base of electrodes or microdialysis probes in chronic experiments. This material allows direct bonding to the cranium.

تطبيق فاتح الشفاء الراتنج لاصق الأسنان أو تركيب لأقطاب مجسات Microdialysis في التجارب المزمنة

In chronic recording experiments, self-curing dental acrylic resins have been used as a mounting base of electrodes or microdialysis-probes. Since these ...

Micro-drive Array for Chronic in vivo Recording: Drive Fabrication

Chronic recording of large populations of neurons is a valuable technique for studying the function of neuronal circuits in awake behaving rats. Lightweight recording devices carrying a high density array of tetrodes allow for the simultaneous monitoring of the activity of tens to hundreds of individual neurons. Here we describe a protocol for the fabrication of a micro-drive array with twenty one independently movable micro-drives. This device has been used successfully to record from hippocampal and cortical neurons in our lab. We show how to prepare a custom designed, 3-D printed plastic base that will hold the micro-drives. We demonstrate how to construct the individual micro-drives and how to assemble the complete micro-drive array. Further preparation of the drive array for surgical implantation, such as the fabrication of tetrodes, loading of tetrodes into the drive array and gold-plating, is covered in a subsequent video article.

Micro-drive Array for Chronic in vivo Recording: Drive Fabrication

In this protocol we demonstrate how to fabricate a micro-drive array for chronic electrophysiological recordings in rats.

الجزئي للمحرك صفيف المزمنة في الجسم الحي تسجيل : تصنيع محرك

Chronic recording of large populations of neurons is a valuable technique for studying the function of neuronal circuits in awake behaving rats. ...

Micro-drive Array for Chronic in vivo Recording: Tetrode Assembly

The tetrode, a bundle of four electrodes, has proven to be a valuable tool for the simultaneous recording of multiple neurons in-vivo.  The differential amplitude of action potential signatures over the channels of a tetrode allows for the isolation of single-unit activity from multi-unit signals.  The ability to precisely control the stereotaxic location and depth of the tetrode is critical for studying coordinated neural activity across brain regions.  In combination with a micro-drive array, it is possible to achieve precise placement and stable control of many tetrodes over the course of days to weeks.  In this protocol, we demonstrate how to fabricate and condition tetrodes using basic tools and materials, install the tetrodes into a multi-drive tetrode array for chronic in-vivo recording in the rat, make ground wire connections to the micro-drive array, and attach a protective cone onto the micro-drive array in order to protect the tetrodes from physical contact with the environment.



Micro-drive Array for Chronic in vivo Recording: Tetrode Assembly

In this protocol we demonstrate how to fabricate and condition tetrodes for use with a micro-drive array, which was designed for chronic electrophysiological recordings in rats. In addition, we illustrate the final stages of micro-drive array construction, which includes installing ground wires and a protective cone.

الجزئي للمحرك صفيف المزمنة في الجسم الحي تسجيل : الجمعية Tetrode

The tetrode, a bundle of four electrodes, has proven to be a valuable tool for the simultaneous recording of multiple neurons in-vivo.  The differential ...

Murine Renal Transplantation Procedure

Renal orthotopic transplantation in mice is a technically challenging procedure. Although the first kidney transplants in mice were performed by Russell et al over 30 years ago (1) and refined by Zhang et al years later (2), few people in the world have mastered this procedure. In our laboratory we have successfully performed 1200 orthotopic kidney transplantations with &gt; 90% survival rate. The key points for success include stringent control of reperfusion injury, bleeding and thrombosis, both during the procedure and post-transplantation, and use of 10-0 instead of 11-0 suture for anastomoses.
Post-operative care and treatment of the recipient is extremely important to transplant success and evaluation. All renal graft recipients receive antibiotics in the form of an injection of penicillin immediately post-transplant and sulfatrim in the drinking water continually. Overall animal health is evaluated daily and whole blood creatinine analyses are performed routinely with a portable I-STAT machine to assess graft function.

Renal transplantation in mice is a technically challenging procedure that requires careful post-operative care and treatment for success.

فأري إجراء زراعة الكلى

Renal orthotopic transplantation in mice is a technically challenging procedure. Although the first kidney transplants in mice were performed by Russell ...

Live Imaging of the Zebrafish Embryonic Brain by Confocal Microscopy

In this video, we demonstrate the method our lab has developed to analyze the cell shape changes and rearrangements required to bend and fold the developing zebrafish brain (Gutzman et al, 2008). Such analysis affords a new understanding of the underlying cell biology required for development of the 3D structure of the vertebrate brain, and significantly increases our ability to study neural tube morphogenesis. The embryonic zebrafish brain is shaped beginning at 18 hours post fertilization (hpf) as the ventricles within the neuroepithelium inflate. By 24 hpf, the initial steps of neural tube morphogenesis are complete. Using the method described here, embryos at the one cell stage are injected with mRNA encoding membrane-targeted green fluorescent protein (memGFP). After injection and incubation, the embryo, now between 18 and 24 hpf, is mounted, inverted, in agarose and imaged by confocal microscopy. Notably, the zebrafish embryo is transparent making it an ideal system for fluorescent imaging. While our analyses have focused on the midbrain-hindbrain boundary and the hindbrain, this method could be extended for analysis of any region in the zebrafish to a depth of 
80-100 &mu;m.

In this video, we demonstrate a method by which to analyze the developing vertebrate brain in live zebrafish embryos at single cell resolution by confocal microscopy. This includes the method by which we inject the single-cell zebrafish embryo and subsequently mount and image the developing brain.

يعيش تصوير الدماغ الجنينية اسماك الزرد بواسطة الميكروسكوب متحد البؤر

In this video, we demonstrate the method our lab has developed to analyze the cell shape changes and rearrangements required to bend and fold the ...

Making MR Imaging Child's Play - Pediatric Neuroimaging Protocol, Guidelines and Procedure

Within the last decade there has been an increase in the use of structural and functional magnetic resonance imaging (fMRI) to investigate the neural basis of human perception, cognition and behavior 1, 2. Moreover, this non-invasive imaging method has grown into a tool for clinicians and researchers to explore typical and atypical brain development. Although advances in neuroimaging tools and techniques are apparent, (f)MRI in young pediatric populations remains relatively infrequent 2. Practical as well as technical challenges when imaging children present clinicians and research teams with a unique set of problems 3, 2. To name just a few, the child participants are challenged by a need for motivation, alertness and cooperation. Anxiety may be an additional factor to be addressed. Researchers or clinicians need to consider time constraints, movement restriction, scanner background noise and unfamiliarity with the MR scanner environment2,4-10. A progressive use of functional and structural neuroimaging in younger age groups, however, could further add to our understanding of brain development. As an example, several research groups are currently working towards early detection of developmental disorders, potentially even before children present associated behavioral characteristics e.g.11. Various strategies and techniques have been reported as a means to ensure comfort and cooperation of young children during neuroimaging sessions. Play therapy 12, behavioral approaches 13, 14,15, 16-18 and simulation 19, the use of mock scanner areas 20,21, basic relaxation 22 and a combination of these techniques 23 have all been shown to improve the participant's compliance and thus MRI data quality. Even more importantly, these strategies have proven to increase the comfort of families and children involved 12. One of the main advances of such techniques for the clinical practice is the possibility of avoiding sedation or general anesthesia (GA) as a way to manage children's compliance during MR imaging sessions 19,20. In the current video report, we present a pediatric neuroimaging protocol with guidelines and procedures that have proven to be successful to date in young children.

Making MR Imaging Child&#039;s Play - Pediatric Neuroimaging Protocol, Guidelines and Procedure

Despite an increase in the use of structural and functional magnetic resonance imaging (fMRI) in humans, the study of young pediatric populations remains a challenge. We present a hands-on, step-by-step video protocol including guidelines for clinicians and researchers intending to perform (f)MRI in young children.

صنع اللعب MR التصوير الطفل -- البروتوكول تصوير الأعصاب للأطفال ، والمبادئ التوجيهية والإجراءات

Within the last decade there has been an increase in the use of structural and functional magnetic resonance imaging (fMRI) to investigate the neural ...

Bioluminescence Imaging of Heme Oxygenase-1 Upregulation in the Gua Sha Procedure

Gua Sha is a traditional Chinese folk therapy that employs skin scraping to cause subcutaneous microvascular blood extravasation and bruises. The protocol for bioluminescent optical imaging of HO-1-luciferase transgenic mice reported in this manuscript provides a rapid in vivo assay of the upregulation of the heme oxygenase-1 (HO-1) gene expression in response to the Gua Sha procedure. HO-1 has long been known to provide cytoprotection against oxidative stress. The upregulation of HO-1, assessed by the bioluminescence output, is thought to represent an antioxidative response to circulating hemoglobin products released by Gua Sha. Gua Sha was administered by repeated strokes of a smooth spoon edge over lubricated skin on the back or other targeted body part of the transgenic mouse until petechiae (splinter hemorrhages) or ecchymosis (bruises) indicative of extravasation of blood from subcutaneous capillaries was observed. After Gua Sha, bioluminescence imaging sessions were carried out daily for several days to follow the dynamics of HO-1 expression in multiple internal organs.

Gua Sha, traditional Chinese therapeutic skin scraping, causes subcutaneous microvascular blood extravasation. We report a protocol of bioluminescence imaging of HO-1-luciferase transgenic mice to demonstrate that Gua Sha upregulates heme oxygenase-1 (HO-1) in multiple organs.

التصوير من تلألؤ بيولوجي Upregulation هيم - 1 أوكسيجيناز في إجراء شا غوا

Gua Sha is a traditional Chinese folk therapy that employs skin scraping to cause subcutaneous microvascular blood extravasation and bruises. The protocol ...

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.

تعيين آثار ما بعد من ثيتا انفجار تحفيز على القشرة السمعية الإنسان مع التصوير الوظيفي

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

Isolation of Human Hepatocytes by a Two-step Collagenase Perfusion Procedure

The liver, an organ with an exceptional regeneration capacity, carries out a wide range of functions, such as detoxification, metabolism and homeostasis. As such, hepatocytes are an important model for a large variety of research questions. In particular, the use of human hepatocytes is especially important in the fields of pharmacokinetics, toxicology, liver regeneration and translational research. Thus, this method presents a modified version of a two-step collagenase perfusion procedure to isolate hepatocytes as described by Seglen 1.
Previously, hepatocytes have been isolated by mechanical methods. However, enzymatic methods have been shown to be superior as hepatocytes retain their structural integrity and function after isolation. This method presented here adapts the method designed previously for rat livers to human liver pieces and results in a large yield of hepatocytes with a viability of 77&plusmn;10%. The main difference in this procedure is the process of cannulization of the blood vessels. Further, the method described here can also be applied to livers from other species with comparable liver or blood vessel sizes.

A modified two-step collagenase perfusion procedure for isolation of human hepatocytes is described. This method can also be applied to other mammalian livers. The isolated hepatocytes are available in high yield and viability, making them a suitable model for scientific research in areas such as liver regeneration, pharmacokinetics and toxicology.

عزل خلايا الكبد البشرية من قبل خطوتين كولاجيناز الإرواء الداخلي

The liver, an organ with an exceptional regeneration capacity, carries out a wide range of functions, such as detoxification, metabolism and homeostasis. ...

Investigating Protein-protein Interactions in Live Cells Using Bioluminescence Resonance Energy Transfer

Assays based on Bioluminescence Resonance Energy Transfer (BRET) provide a sensitive and reliable means to monitor protein-protein interactions in live cells. BRET is the non-radiative transfer of energy from a 'donor' luciferase enzyme to an 'acceptor' fluorescent protein. In the most common configuration of this assay, the donor is Renilla reniformis luciferase and the acceptor is Yellow Fluorescent Protein (YFP). Because the efficiency of energy transfer is strongly distance-dependent, observation of the BRET phenomenon requires that the donor and acceptor be in close proximity. To test for an interaction between two proteins of interest in cultured mammalian cells, one protein is expressed as a fusion with luciferase and the second as a fusion with YFP. An interaction between the two proteins of interest may bring the donor and acceptor sufficiently close for energy transfer to occur. Compared to other techniques for investigating protein-protein interactions, the BRET assay is sensitive, requires little hands-on time and few reagents, and is able to detect interactions which are weak, transient, or dependent on the biochemical environment found within a live cell. It is therefore an ideal approach for confirming putative interactions suggested by yeast two-hybrid or mass spectrometry proteomics studies, and in addition it is well-suited for mapping interacting regions, assessing the effect of post-translational modifications on protein-protein interactions, and evaluating the impact of mutations identified in patient DNA.

Interactions between proteins are fundamental to all cellular processes. Using Bioluminescence Resonance Energy Transfer, the interaction between a pair of proteins can be monitored in live cells and in real time. Furthermore, the effects of potentially pathogenic mutations can be assessed.

التحقيق البروتين البروتين التفاعلات في الخلايا الحية باستخدام الرنين ضوء بارد نقل الطاقة

Assays based on Bioluminescence Resonance Energy Transfer (BRET) provide a sensitive and reliable means to monitor protein-protein interactions in live ...