Name: a large lateral craniotomy procedure for mesoscale wide-field optical imaging of brain activity
Uploaded: 2017-05-07T13:00:00
Description: Watch this Scientific Journal Video about A Large Lateral Craniotomy Procedure for Mesoscale Wide-field Optical Imaging of Brain Activity at JoVE.com

Эта работа была поддержана естественных и технических наук исследовательский совет Канады (NSERC) Discovery Grant # 40352, Campus Alberta инновационной программы кафедры, программы исследований Альберты Альцгеймера в MHM, и NSERC CREATE в БИФ докторских стипендий и AIHS послевузовского общения с МК. Мы благодарим Pu Мин Ван для развития этого протокола и для хирургической подготовки и Беру Мирза Ага и Ди Шао для животноводства.

<html> Следующий протокол следует Университет Летбридж животных комитета Care (ACC) руководящих принципов, и проводится в соответствии со стандартами Канадского совета по уходу за животными (ССАС). 1. Подготовка <ol><li> Для длительных периодов исследования, автоклав всех открыт?...

<ol>
	<li>Sigler, A., Mohajerani, M. H., Murphy, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+rapid+redistribution+of+sensory-evoked+depolarization+through+existing+cortical+pathways+after+targeted+stroke+in+mice.">Imaging rapid redistribution of sensory-evoked depolarization through existing cortical pathways after targeted stroke in mice.</a> Proc Natl Acad Sci U S A. 106 (28), 11759-11764 (2009).</li><li>Shih, A. Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two-photon+microscopy+as+a+tool+to+study+blood+flow+and+neurovascular+coupling+in+the+rodent+brain.">Two-photon microscopy as a tool to study blood flow and neurovascular coupling in the rodent brain.</a> J Cereb Blood Flow Metab. 32 (7), 1277-1309 (2012).</li><li>Grinvald, A., Hildesheim, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=VSDI:+a+new+era+in+functional+imaging+of+cortical+dynamics.">VSDI: a new era in functional imaging of cortical dynamics.</a> Nat Rev Neurosci. 5 (11), 874-885 (2004).</li><li>Blinder, P., Shih, A. Y., Rafie, C., Kleinfeld, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Topological+basis+for+the+robust+distribution+of+blood+to+rodent+neocortex.">Topological basis for the robust distribution of blood to rodent neocortex.</a> Proc Natl Acad Sci U S A. 107 (28), 12670-12675 (2010).</li><li>Mohajerani, M. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spontaneous+cortical+activity+alternates+between+motifs+defined+by+regional+axonal+projections.">Spontaneous cortical activity alternates between motifs defined by regional axonal projections.</a> Nat Neurosci. 16 (10), 1426-1435 (2013).</li><li>Mohajerani, M. H., McVea, D. A., Fingas, M., Murphy, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mirrored+bilateral+slow-wave+cortical+activity+within+local+circuits+revealed+by+fast+bihemispheric+voltage-sensitive+dye+imaging+in+anesthetized+and+awake+mice.">Mirrored bilateral slow-wave cortical activity within local circuits revealed by fast bihemispheric voltage-sensitive dye imaging in anesthetized and awake mice.</a> J Neurosci. 30 (10), 3745-3751 (2010).</li><li>Lippert, M. T., Takagaki, K., Xu, W., Huang, X., Wu, J. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+for+voltage-sensitive+dye+imaging+of+rat+cortical+activity+with+high+signal-to-noise+ratio.">Methods for voltage-sensitive dye imaging of rat cortical activity with high signal-to-noise ratio.</a> J Neurophysiol. 98 (1), 502-512 (2007).</li><li>Misgeld, T., Kerschensteiner, M. <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+imaging+of+the+diseased+nervous+system.">In vivo imaging of the diseased nervous system.</a> Nat Rev Neurosci. 7 (6), 449-463 (2006).</li><li>Kerr, J. N., Denk, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+in+vivo:+watching+the+brain+in+action.">Imaging in vivo: watching the brain in action.</a> Nat Rev Neurosci. 9 (3), 195-205 (2008).</li><li>Aronoff, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-range+connectivity+of+mouse+primary+somatosensory+barrel+cortex.">Long-range connectivity of mouse primary somatosensory barrel cortex.</a> Eur J Neurosci. 31 (12), 2221-2233 (2010).</li><li>McVea, D. A., Mohajerani, M. H., Murphy, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Voltage-sensitive+dye+imaging+reveals+dynamic+spatiotemporal+properties+of+cortical+activity+after+spontaneous+muscle+twitches+in+the+newborn+rat.">Voltage-sensitive dye imaging reveals dynamic spatiotemporal properties of cortical activity after spontaneous muscle twitches in the newborn rat.</a> J Neurosci. 32 (32), 10982-10994 (2012).</li><li>Sweetnam, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diabetes+impairs+cortical+plasticity+and+functional+recovery+following+ischemic+stroke.">Diabetes impairs cortical plasticity and functional recovery following ischemic stroke.</a> J Neurosci. 32 (15), 5132-5143 (2012).</li><li>Yin, Y. Q., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+field+recordings+effectively+monitor+the+mouse+cortex+and+hippocampus+under+isoflurane+anesthesia.">In vivo field recordings effectively monitor the mouse cortex and hippocampus under isoflurane anesthesia.</a> Neural Regeneration Research. 11 (12), 1951-1955 (2016).</li><li>Sharp, P. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+stimulus-evoked+cerebral+hemodynamics+in+the+awake+mouse+and+under+a+novel+anesthetic+regime.">Comparison of stimulus-evoked cerebral hemodynamics in the awake mouse and under a novel anesthetic regime.</a> Scientific Reports. 5, 12621 (2015).</li><li>Kyweriga, M., Mohajerani, M. H., Kianianmomeni, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optogenetics:+Methods+and+Protocols.">Optogenetics: Methods and Protocols.</a> Methods in Molecular Biology. 1408, 251-265 (2016).</li><li>Grutzendler, J., Gan, W. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Imaging in neuroscience and development : a laboratory manual. , (2005).</li><li>Vanni, M. P., Murphy, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mesoscale+transcranial+spontaneous+activity+mapping+in+GCaMP3+transgenic+mice+reveals+extensive+reciprocal+connections+between+areas+of+somatomotor+cortex.">Mesoscale transcranial spontaneous activity mapping in GCaMP3 transgenic mice reveals extensive reciprocal connections between areas of somatomotor cortex.</a> J Neurosci. 34 (48), 15931-15946 (2014).</li><li>Xie, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Resolution+of+High-Frequency+Mesoscale+Intracortical+Maps+Using+the+Genetically+Encoded+Glutamate+Sensor+iGluSnFR.">Resolution of High-Frequency Mesoscale Intracortical Maps Using the Genetically Encoded Glutamate Sensor iGluSnFR.</a> J Neurosci. 36 (4), 1261-1272 (2016).</li><li>Chan, A. W., Mohajerani, M. H., LeDue, J. M., Wang, Y. T., Murphy, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mesoscale+infraslow+spontaneous+membrane+potential+fluctuations+recapitulate+high-frequency+activity+cortical+motifs.">Mesoscale infraslow spontaneous membrane potential fluctuations recapitulate high-frequency activity cortical motifs.</a> Nat Commun. 6, 7738 (2015).</li><li>Lim, D. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+Large-Scale+Cortical+Mapping+Using+Channelrhodopsin-2+Stimulation+in+Transgenic+Mice+Reveals+Asymmetric+and+Reciprocal+Relationships+between+Cortical+Areas.">In vivo Large-Scale Cortical Mapping Using Channelrhodopsin-2 Stimulation in Transgenic Mice Reveals Asymmetric and Reciprocal Relationships between Cortical Areas.</a> Front Neural Circuits. 6, (2012).</li><li>Ferezou, I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatiotemporal+dynamics+of+cortical+sensorimotor+integration+in+behaving+mice.">Spatiotemporal dynamics of cortical sensorimotor integration in behaving mice.</a> Neuron. 56 (5), 907-923 (2007).</li><li>Mohajerani, M. H., Aminoltejari, K., Murphy, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeted+mini-strokes+produce+changes+in+interhemispheric+sensory+signal+processing+that+are+indicative+of+disinhibition+within+minutes.">Targeted mini-strokes produce changes in interhemispheric sensory signal processing that are indicative of disinhibition within minutes.</a> Proc Natl Acad Sci U S A. 108 (22), E183-E191 (2011).</li><li>Madisen, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transgenic+mice+for+intersectional+targeting+of+neural+sensors+and+effectors+with+high+specificity+and+performance.">Transgenic mice for intersectional targeting of neural sensors and effectors with high specificity and performance.</a> Neuron. 85 (5), 942-958 (2015).</li></ol>

Этот инновационный протокол для большого черепного окна позволяет одновременно томографию над височной и теменной областей коры головного мозга. В сочетании с оптической визуализацией, он может помочь выявить нейронную динамику в пределах областей коры при спонтанной и стимуле-инду...

Краниотомию является стандартной процедурой используется неврологами, чтобы показать часть мозга. Испокон электрофизиологии, трепанация позволила беспрецедентным прорывы в области нейробиологии. Плотные картирование коры головного мозга с электродами приводят к экспериментам для проверки гипотез и теорий, основанных на этих картах. Недавно мы вступили в новую эру , где краниотомия утилизируется для получения изображения в естественных условиях коркового кровотока 1, 2, 3 и сосудисто архитектуры 4, что позволяет в реальном времени визуализации корковой активности в открытых участках 5, 6, 7. Хотя многие исследования используют краниотомии в сочетании с методами оптической визуализации в естественных условиях для изучения структуры и функции корковых нейронов, глии и кора8 , 9 дальнейшие исследования ограничены небольшими участками открытой коры (см. 10 ). Цель этого протокола заключается в том, чтобы предоставить способ создания крупной боковой краниотомии, подвергая кору головного мозга от средней линии до кости чешуи и выходящей за пределы брегмы и лямбды. Эта крупная краниотомия позволяет одновременно наблюдать коры ассоциации (ретросплениальный, поясной и париетальный), первичный и вторичный мотор, соматосенсорный, зрительный и слуховой коры. Этот метод ранее был связан с чувствительным к напряжению изображением красителя (VSDI), чтобы исследовать, как несколько областей коры взаимодействуют друг с другом во время спонтанной и индуцированной стимулом кортикальной активности 5 , 11 , 12 . Наиболее сложными аспектами этой процедуры являются позиционирование головыживотное, фиксируя пластины головки, и избегая кровоизлияние при отделении височной мышцы от теменной кости. Кроме того, следует принимать во время процессов бурения и удаления черепа как кривые черепов под косым углом.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Heating Pad&#160;</td>
			<td>FHC</td>
			<td>40-90-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Fine Scissors</td>
			<td>Fine Science Tools</td>
			<td>14058-09</td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps&#160;</td>
			<td>Fine Science Tools</td>
			<td>11251-35</td>
			<td>2 or more pairs are recommended</td>
		</tr>
		<tr>
			<td>Spring scissors</td>
			<td>Fine Science Tools</td>
			<td>15000-00, 15000-10</td>
			<td>1 pair should be designated for dura removal&#160;</td>
		</tr>
		<tr>
			<td>Jet tooth shade powder</td>
			<td>LANG Dental</td>
			<td>Jet Tooth Shade Powder</td>
			<td>to be mixed with the Jet Liquid</td>
		</tr>
		<tr>
			<td>Jet tooth shade liquid</td>
			<td>LANG Dental</td>
			<td>Jet Tooth Shade Liquid</td>
			<td>to be mixed wihth the Jet Powder&#160;</td>
		</tr>
		<tr>
			<td>Drill Heads - Carbide Burs FG 1/4 389</td>
			<td>Midwest Dental</td>
			<td>385201</td>
			<td></td>
		</tr>
		<tr>
			<td>Agarose Powder</td>
			<td>Sigma-Aldrich</td>
			<td>A9793</td>
			<td></td>
		</tr>
		<tr>
			<td>Gelfoam</td>
			<td>Sinclair Dental Canada</td>
			<td>Pfizer Gelfoam</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Western Drug Distribution Centre Ltd</td>
			<td>124125</td>
			<td></td>
		</tr>
		<tr>
			<td>Lidocaine 2% Epinephrine</td>
			<td>Western Drug Distribution Centre Ltd</td>
			<td>125299</td>
			<td></td>
		</tr>
		<tr>
			<td>Dexamethazone 5 mg/mL</td>
			<td>Western Drug Distribution Centre Ltd</td>
			<td>125231</td>
			<td></td>
		</tr>
		<tr>
			<td>Butyl cyanoacrylate glue (VetBond)</td>
			<td>Western Drug Distribution Centre Ltd</td>
			<td>12612</td>
			<td></td>
		</tr>
	</tbody>
</table>

a large lateral craniotomy procedure for mesoscale wide-field optical imaging of brain activity

Краниотомия - это обычно выполняемая процедура, при которой мозг подвергается экспериментам in vivo . В исследованиях мыши большинство лабораторий используют небольшую краниотомию, обычно 3 мм x 3 мм. Этот протокол вводит метод создания значительно большего 7 мм х 6 мм краниального окна, обнажающего большую часть полушария головного мозга над височной и теменной корой ( например, bregma 2,5-4,5 мм, боковые 0-6 мм). Чтобы выполнить эту операцию, голова должна быть наклонена приблизительно на 30 °, и большая часть височной мышцы должна быть втянута. Из-за большого количества удаления кости эта процедура предназначена только для острых экспериментов с животными, которых анестезировали во время операции и эксперимента. Главным преимуществом этого инновационного большого бокового краниального окна является одновременный доступ как к медиальной, так и к боковым областям коры. Это большое одностороннее краниальное окно может быть использовано для изучения нейронной динамики между клетками,а также между различными областями коры путем объединения нескольких электродов электрофизиологических записей, визуализации нейронной активности (например, внутреннее или внешнее изображения), и оптогенетики стимуляции. Кроме того, эта большая краниотомия также предоставляет большую площадь корковых кровеносных сосудов, что позволяет для прямого манипулирования бокового корковых сосудистой сети.

<html> Для изучения взаимодействий между областями коры в пределах одного полушария, мы использовали большую краниотомию, проходящую через сагиттальный синус и 5 - 6 мм сбоку. Это окно черепных входит первичный (двигатель, соматосенсорной, зрительные, слуховые), вторичный ...

Watch this Scientific Journal Video about A Large Lateral Craniotomy Procedure for Mesoscale Wide-field Optical Imaging of Brain Activity at JoVE.com

A Large Lateral Craniotomy Procedure for Mesoscale Wide-field Optical Imaging of Brain Activity

Этот протокол представляет собой метод создания большого односторонний краниотомия над височной и теменной областей коры головного мозга мыши. Это особенно полезно для реального времени визуализации более экспансивной области коркового полушария.

A Большой процедура Боковой Краниотомии для мезомасштабного широкоугольных оптического Визуализация активности мозга

The craniotomy is a commonly performed procedure to expose the brain for in vivo experiments. In mouse research, most labs utilize a small craniotomy, ...

The overall goal of this large lateral craniotomy is to perform real time imaging over an expansive area of a cortical hemisphere. The main advantage of this technique is that large areas of the cortex are exposed for observation and can be used in conjunction with technologies such as optogenetics. This technique can provide insights into characterizing the neural dynamics between cells as well as that between different brain circuits at different temporal and spatial resolutions.Visual demonstration of this method is critical due to the increased risk of hemmorage and cortical damage since the dura is often attached to brain surface and blood vessels. This method can be applied to study the neurobiological mechanisms underlying brain circuit development and its changes throughout life in response to new experiences or pathologies such as Alzheimer's Disease or stroke. To begin this procedure, autoclave all the surgical supplies and ensure that sterility is maintained throughout the surgery.Ensure there is enough brain buffer for the surgery. Next, transfer an anesthetized mouse to the head holder setup and place it on a thermo-regulating heating pad set to 37 degrees Celsius. Secure the mouse's upper teeth in a teeth holder.Then rotate its head towards the left approximately 30 degrees to expose the right lateral side of the head. Subsequently, secure the mouse's head with the blunt end of the ear bars. Then apply a thalmic ointment to prevent corneal drying.To reduce cerebral edema, inject it with dexamethasone at four milligrams per kilogram intermuscularly. Wipe the skin over the surgical area with cotton swabs dipped in 4%chlorhexidine three times and then with 70%alcohol three times. Inject lidocaine subcutaneously over the craniotomy site.Under a dissecting microscope, lift up the skin at one millimeter left of the midline just behind the ear with forceps and make a small horizontal incision with surgical scissors. Make a five to six millimeter lateral cut towards the right ear and then cut towards the rostral end of the head. At the initial incision point, insert the scissors and cut 10 millimeters rostrally.After that, cut the skin around the right ear and near the right eye to expose the right side of the skull and temporal muscle. Ensure that the widest part of the exposed area is at least seven millimeters. Then trim the skin further if the surgical area needs to be extended.Using a cotton swab, rub the surface of the skull in a circular motion to remove the periosteum from the skull. Ensure that no periosteum remains and that the skull is completely dry, which is critical for successful attachment of the head plate. Next, using spring scissors and forceps, separate the temporal muscle from the skull.Cut and retract the muscle laterally until it reaches the squamosal bone. Take extreme care not to damage the superficial temporal vein that runs along the level of the squamosal bone near the eye, otherwise hemorrhage might occur. Control bleeding with gel foam pre-soaked in brain buffer.For serious hemorrhage, use a heat cauterizer. Drop butyl cyanoacrylate glue onto bleeding sites. To begin this procedure, first mark bregma to orient the underlying brain regions, apply ethyl cyanoacrylate glue around the bottom edge of the head plate and glue the head plate over the craniotomy area.After that, fill the opening space between the skull and the head plate with dental cement, leaving only the craniotomy area exposed. Then wait for the dental cement to dry and harden. Subsequently, outline the cranial window region by lightly scoring the surface of the skull with the dental drill.Gently trace the drill along the original scoring to deepen it, ensuring that the drill does not penetrate through the skull into the brain. In the meantime, switch between drilling and dabbing the skull's surface with moistened rolled tissues every few minutes. This very challenging start requires surgeons to drill through the aged skull to free it for removal without damaging the underlying cortex.Slow is loose, smooth is fast. During drilling, periodically check for buckling of the skull by gently pressing on it with forceps or the non-moving drill bit. When the bone begins the buckle, stop drilling and immerse the entire window in brain buffer.Wait for at least five minutes before skull removal to soften the bone and to reduce the chance of the dura sticking to the bone, making the skull removal process easier. Next, perform the skull removal process beginning at the anterior edge. Gently pry the loose skull from the dura using forceps.Once the bone is loose and floating on the dura, firmly grasp it with forceps and lift it away from the dura. Ensure the bone never penetrates into the brain. Use forceps and spring scissors to gently tear and cut away the dura.Care must be taken to ensure the brain parenchyma and blood vessels are not damaged. Continue until all dura is removed from the cranial window site. When performed correctly, the brain will appear to be very clean with distinct blood vessels and no blemishes.Spray the coverglass with 70%ethanol and use an air canister to gently dry. Ensure the glass is completely clean with no spots or dust present. Next, prepare 1.3%agarose by heating 200 milligrams of agarose powder dissolved in 15 milliliters of brain buffer.Place a thermometer in the hot agarose and cool the agarose down to just above solidifying temperature at about 40 degrees Celsius. Cooling can be expedited by running cold water over the tube, taking care not to allow any water to enter the agar. Gently stir with the thermometer while monitoring its temperature.Immediately before applying the agar, remove the brain buffer from the cranial well. Ensure no bubbles or particles are present as these will interfere with imaging. Now draw up the agarose with a transfer pipe head and drop the agarose directly on the brain.Then quickly place the coverslip over the surface and fasten the coverslip with the agarose drops on the corners. Shown here is a photo micrograph of the wide unilateral craniotomy with bregma indicated by a white circle in each image. Patterns of cortical activation are shown in a mouse anesthetized with 0.5%isoflurane after stimulation of the contralateral whisker, auditory stimulation, contralateral forelimb stimulation, contralateral hindlimb stimulation and visual stimulation of the contralateral eye with a light-emitting diode.There was midline activation from all forms of sensory stimulation at 10 to 25 milliseconds after primary sensory cortex activation. Once mastered, this technique can be completed in three to four hours. When attempting this procedure, it's important to remember to keep the blood loss to minimum and to ensure the cortex is not damaged.Using this procedure, other imaging techniques such as optical imaging can help to answer important questions such as how neural dynamics are influenced by spontaneous activity in the normal and diseased brain. Following its development, this technique helped to pave the way for researchers in the field of systems neuroscience to explore interactions and neural dynamics between widely spaced cortical regions in mice. This video shows you how to perform a large lateral craniotomy in the mouse with minimal blood loss and cortical damage, enabling you to collect the necessary data to answer your most pressing scientific questions.

Research

JoVE Journal

Neuroscience

State-Dependency Effects on TMS:  A Look at Motive Phosphene Behavior

Государство-зависимость Влияние на TMS: Посмотрите на мотив поведения фосфен

Transcranial magnetic stimulation (TMS) is a non-invasive neurostimulatory and neuromodulatory technique that can transiently or lastingly modulate ...

Live Imaging of the Ependymal Cilia in the Lateral Ventricles of the Mouse Brain

Живая съемка в эпендимных ресничек боковых желудочков головного мозга мыши

Multiciliated ependymal cells line the ventricles in the adult brain. Abnormal function or structure of ependymal cilia is associated with various ...

Modification of a Colliculo-thalamocortical Mouse Brain Slice, Incorporating 3-D printing of Chamber Components and Multi-scale Optical Imaging

Модификация Colliculo-таламокортикального мозга мыши фрагмент, включающий 3-D печать палаты компонентов и нескольких масштаб оптических изображений

The ability of the brain to process sensory information relies on both ascending and descending sets of projections. Until recently, the only way to study ...

Large-scale Three-dimensional Imaging of Cellular Organization in the Mouse Neocortex

Крупномасштабные трехмерных изображений клеточной организации коры головного мозга мыши

The mammalian neocortex is composed of many types of excitatory and inhibitory neurons, each with specific electrophysiological and biochemical ...

Wide-field Single-photon Optical Recording in Brain Slices Using Voltage-sensitive Dye

Широкоугольной однофотонной оптической записи в мозговых фрагментах с использованием чувствительных к напряжению красителя

Wide-field single photon voltage-sensitive dye (VSD) imaging of brain slice preparations is a useful tool to assess the functional connectivity in neural ...

Real-Time fMRI Brain Mapping in Animals

Картирование мозга фМРТ в реальном времени у животных

Dynamic fMRI responses vary largely according to the physiological conditions of animals either under anesthesia or in awake states. We developed a ...

Activity of Posterior Lateral Line Afferent Neurons during Swimming in Zebrafish

Активность афферентных нейронов задней боковой линии во время плавания у рыбок данио

Sensory systems gather cues essential for directing behavior, but animals must decipher what information is biologically relevant. Locomotion generates ...

Craniotomy Procedure for Visualizing Neuronal Activities in Hippocampus of Behaving Mice

Процедура краниотомии для визуализации нейронной активности в гиппокампе мышей

Imaging neuronal activities at single-cell resolution in awake behaving animals is a very powerful approach for the investigation of neural circuit ...

In Vivo Wide-Field and Two-Photon Calcium Imaging from a Mouse Using a Large Cranial Window

In Vivo Wide-Field and Two-Photon Calcium Imaging from a Mouse Using a Large Cranial Window

В городе Виво Широкоугольная и двухфотонная визуализация кальция с помощью мыши с помощью большого краниального окна

Wide-field calcium imaging from the mouse&#39;s neocortex allows one to observe cortex-wide neural activity related to various brain functions. On the ...

Recording Brain Activity with Ear-Electroencephalography

Запись активности мозга с помощью электроэнцефалографии уха

The c-grid (ear-electroencephalography, sold under the name cEEGrid) is an unobtrusive and comfortable electrode array that can be used for investigating ...