Name: exposure of the pig cns for histological analysis: a manual for decapitation, skull opening, and brain removal
Uploaded: 2017-04-13T14:00:00
Description: Watch this Scientific Journal Video about Exposure of the Pig CNS for Histological Analysis: A Manual for Decapitation, Skull Opening, and Brain Removal at JoVE.com

The authors acknowledge with gratitude the skillful assistance of Mrs. Trine W. Mikkelsen, Mrs. Lise M. Fitting, and the staff at P&#229;skeh&#248;jgaard. The Danish Medical Research Council, the Lundbeck Foundation, and the Novo Nordisk Foundation financially supported the study.

Animal anesthesia and euthanesia was performed in accordance with &#34;Principles of laboratory animal care&#34; (NIH publication No. 86-23, revised 1985) and approved by the Danish Council for Animal Research Ethics.
1. Instruments
<ol>
	<li>Collect the instruments presented in the video and listed in the Table of Materials.</li>
</ol>
2. Decapitation
NOTE: Anesthesia was induced by an intramuscular injection of 5 mL of mida...

<ol>
	<li>Lind, M. N., Moustgaard, A., Jelsing, J., Vajta, G., Cumming, P., Hansen, A. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+pigs+in+neuroscience:+Modeling+brain+disorders.">The use of pigs in neuroscience: Modeling brain disorders.</a> Neurosci Biobehav Rev. 31, 728-751 (2007).</li><li>Bjarkam, C. 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=Neuromodulation+in+a+minipig+model+of+Parkinson+disease.">Neuromodulation in a minipig model of Parkinson disease.</a> British J Neurosurg. 22 (Suppl. 1), S9-S12 (2008).</li><li>Bjarkam, C. R., Cancian, G., Glud, A. N., Ettrup, K. S., &#216;stergaard, L., S&#248;rensen, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MRI-guided+stereotaxic+targeting+in+pigs+based+on+a+stereotaxic+localizer+box+fitted+with+an+isocentric+frame+and+use+of+SurgiPlan+computer-planning+software.">MRI-guided stereotaxic targeting in pigs based on a stereotaxic localizer box fitted with an isocentric frame and use of SurgiPlan computer-planning software.</a> J Neurosci Methods. 183 (2), 119-126 (2009).</li><li>Sauleau, P., Lapouble, E., Val-Laillet, D., Malbert, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+pig+model+in+brain+imaging+and+neurosurgery.">The pig model in brain imaging and neurosurgery.</a> Animal. 3 (8), 1138-1151 (2009).</li><li>Bjarkam, C. 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=Safety+and+function+of+a+new+clinical+intracerebral+microinjection+instrument+for+stem+cells+and+therapeutics+examined+in+the+Göttingen+minipig.">Safety and function of a new clinical intracerebral microinjection instrument for stem cells and therapeutics examined in the Göttingen minipig.</a> Stereotact Funct Neurosurg. 88 (1), 56-63 (2010).</li><li>Fjord-Larsen, 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=Long-term+delivery+of+nerve+growth+factor+by+encapsulated+cell+biodelivery+in+the+minipig+basal+forebrain.">Long-term delivery of nerve growth factor by encapsulated cell biodelivery in the minipig basal forebrain.</a> Mol Therapy. 18 (12), 2164-2172 (2010).</li><li>S&#248;rensen, J. C., 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=Development+of+neuromodulation+treatments+in+a+large+animal+model+-+Do+neurosurgeons+dream+of+electric+pigs?.">Development of neuromodulation treatments in a large animal model - Do neurosurgeons dream of electric pigs?.</a> Prog Brain Res. 194, 97-103 (2011).</li><li>Dolezalova, 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=Pig+models+of+neurodegenerative+disorders:+utilization+in+cell+replacement-based+preclinical+safety+and+efficacy+studies.">Pig models of neurodegenerative disorders: utilization in cell replacement-based preclinical safety and efficacy studies.</a> J Comp Neurol. 522 (12), 2784-2801 (2014).</li><li>Mikkelsen, M., Moller, A., Jensen, L. H., Pedersen, A., Harajehi, J. 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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=The+DaNEX+study+of+embryonic+mesencephalic,+dopaminergic+tissue+grafted+to+a+minipig+model+of+Parkinson's+disease:+Preliminary+findings+of+effect+of+MPTP+poisoning+on+striatal+dopaminergic+markers.">The DaNEX study of embryonic mesencephalic, dopaminergic tissue grafted to a minipig model of Parkinson's disease: Preliminary findings of effect of MPTP poisoning on striatal dopaminergic markers.</a> Cell Transplant. 9 (2), 247-259 (2000).</li><li>Dalmose, A., Bjarkam, C. R., S&#248;rensen, J. C., J&#248;rgensen, T. M., Djurhuus, J. 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H., Cumming, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pig+brain+stereotaxic+standard+space:+Mapping+of+cerebral+blood+flow+normative+values+and+effect+of+MPTP-lesioning.">Pig brain stereotaxic standard space: Mapping of cerebral blood flow normative values and effect of MPTP-lesioning.</a> Brain Res Bull. 66 (1), 17-29 (2005).</li><li>Glud, A. N., 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=Direct+gene+transfer+in+the+minipig+CNS+using+stereotaxic+lentiviral+microinjections.">Direct gene transfer in the minipig CNS using stereotaxic lentiviral microinjections.</a> Acta Neurobiol Exp. 70 (3), 1-8 (2010).</li><li>Glud, A. N., 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=Direct+MRI-guided+stereotaxic+viral+mediated+gene+transfer+of+alpha-synuclein+in+the+minipig+CNS.">Direct MRI-guided stereotaxic viral mediated gene transfer of alpha-synuclein in the minipig CNS.</a> Acta Neurobiol Exp. 71 (4), 508-518 (2011).</li><li>Ettrup, K. S., S&#248;rensen, J. C., Rodell, A., Alstrup, A. K. O., Bjarkam, C. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypothalamic+deep+brain+stimulation+influences+autonomic+and+limbic+circuitry+involved+in+the+regulation+of+aggression+and+cardiocerebrovascular+control+in+the+minipig.">Hypothalamic deep brain stimulation influences autonomic and limbic circuitry involved in the regulation of aggression and cardiocerebrovascular control in the minipig.</a> Stereotact Funct Neurosurg. 90 (5), 281-291 (2012).</li><li>Nielsen, M. 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=Continuous+MPTP+intoxication+in+the+minipig+results+in+chronic+parkinsonian+deficits.">Continuous MPTP intoxication in the minipig results in chronic parkinsonian deficits.</a> Acta Neurobiol Exp. 76, 198-210 (2016).</li><li>Bjarkam, C. 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=A+MRI-compatible+stereotaxic+localizer+box+enables+high-precision+stereotaxic+procedures+in+pigs.">A MRI-compatible stereotaxic localizer box enables high-precision stereotaxic procedures in pigs.</a> J Neurosci Methods. 139 (2), 293-298 (2004).</li><li>Bjarkam, C. R., Jorgensen, R. L., Jensen, K. N., Sunde, N. A. A., S&#248;rensen, J. C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deep+brain+stimulation+electrode+anchoring+using+BioGlue&#174;,+a+protective+electrode+covering,+and+a+titanium+microplate.">Deep brain stimulation electrode anchoring using BioGlue&#174;, a protective electrode covering, and a titanium microplate.</a> J Neurosci Methods. 168, 151-155 (2008).</li><li>Ettrup, K. 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=Basic+Surgical+Techniques+in+the+Minipig:+Intubation,+Transurethral+Bladder+Catheterization,+Femoral+Vessel+Catheterization,+and+Transcardial+Perfusion.">Basic Surgical Techniques in the Minipig: Intubation, Transurethral Bladder Catheterization, Femoral Vessel Catheterization, and Transcardial Perfusion.</a> J Vis Exp. (52), e2652 (2011).</li><li>Ettrup, K. S., Torn&#248;e, J., S&#248;rensen, J. C., Bjarkam, C. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+surgical+device+for+minimally+invasive+implantation+of+experimental+deep+brain+stimulation+leads+in+large+research+animals.">A surgical device for minimally invasive implantation of experimental deep brain stimulation leads in large research animals.</a> J Neurosci Methods. 200 (1), 41-46 (2011).</li><li>Danielsen, E. 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=Positron+emission+tomography+of+living+brain+in+minipigs+and+domestic+pigs.">Positron emission tomography of living brain in minipigs and domestic pigs.</a> Scand J Lab Anim Sci Suppl. 25 (1), 127-135 (1998).</li><li>R&#248;hl, 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=Time+evolution+of+cerebral+perfusion+and+ADC+measured+by+MRI+in+a+porcine+stroke+model.">Time evolution of cerebral perfusion and ADC measured by MRI in a porcine stroke model.</a> J Magn Reson Imaging. 15 (2), 123-129 (2002).</li><li>Cumming, P., Gillings, N. M., Jensen, S. B., Bjarkam, C. 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R. <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+implantation+of+deep+brain+stimulation+electrodes+in+the+pontine+micturition+centre+of+the+minipig.">Long-term implantation of deep brain stimulation electrodes in the pontine micturition centre of the minipig.</a> Acta Neurochir. 151 (7), 785-794 (2009).</li><li>Rosendal, F., 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=Does+chronic+low+dose+treatment+with+ciclosporin+influence+the+brain?+A+histopathological+study+in+pigs.">Does chronic low dose treatment with ciclosporin influence the brain? A histopathological study in pigs.</a> Transplantation Proc. 37 (8), 3305-3308 (2005).</li><li>Nielsen, M. S., S&#248;rensen, J. C., Bjarkam, C. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+substantia+nigra+pars+compacta+of+the+minipig:+An+anatomical+and+stereological+study.">The substantia nigra pars compacta of the minipig: An anatomical and stereological study.</a> Brain Struct Funct. (4-5), 481-488 (2009).</li><li>S&#248;rensen, J. C., Bjarkam, C. R., Simonsen, C. Z., Danielsen, E., Geneser, F. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oriented+sectioning+of+irregular+tissue+blocks+in+relation+to+computerized+scanning+modalities.+Results+from+the+domestic+pig+brain.">Oriented sectioning of irregular tissue blocks in relation to computerized scanning modalities. Results from the domestic pig brain.</a> J Neurosci Methods. 104, 93-98 (2000).</li><li>Bjarkam, C. R., Pedersen, M., S&#248;rensen, J. 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Most experimental neuroscience studies are performed in small animal species, such as mice and rats, where access to the CNS is facilitated by a thin skull- and dural-thickness. However, in larger experimental animals like pigs1,4,8, sheep32, and non-human primates, the considerable thickness of these structures necessitates the use of robust instruments (table of materials) and proper entry points for sk...

Translational neuroscience studies in pigs have become increasingly popular during the last two decades. The large size of the pig brain enables the use of conventional clinical brain imagers and the direct use and testing of neurosurgical procedures and equipment from the human clinic1,2,3,4,5,6,7,8. In the last 20 years, pigs, especially minipigs (e.g., G&#246;ttingen minipig), have been used to examine neuromodulatory treatment modalities, such as stem cell transplantation; viral vector transfection; and deep brain stimulation directed towards Parkinson disease, obesity, depression, and Alzheimer disease2,6,9,10,11,12,13,14,15,16,17. This has been followed by the development of stereotaxic and surgical approaches to manipulate the minipig CNS3,18,19,20,21. The instituted CNS changes have been evaluated in live animals using brain imaging (PET10,13,22,24 and MR23), cystometry11,12,25, gait analysis17, neurological evaluation9,17, and postmortem examination based on histology and stereological analysis14,15,17,26,27,31. However, postmortem analysis requires the exposure and removal of the pig brain, which is not an easy task, as a thick, bony skull and a fibrous dural covering surround the pig brain.
The goal of this paper and instructional video is to describe how the postmortem pig brain and pituitary may be exposed and removed in an intact state in 15-20 min using non-motorized surgical tools. The instructional video and photographic illustrations show male minipigs (age: 6 months, bodyweight: 20-25 kg) used for an anatomical study on the minipig pituitary gland.

<table border="0" cellpadding="0" cellspacing="0" width="1036">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Heavy Scalpel Handle #4</td>
			<td style="width:191px;">FST (Fine Science Tools)</td>
			<td style="width:149px;">10008-13</td>
			<td style="width:481px;">Good for skin incision and soft tissue removal</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Non-Sterile Scalpel Blades #23</td>
			<td style="width:191px;">FST&#160;</td>
			<td>10023-00</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Scalpel Handle #7</td>
			<td style="width:191px;">FST&#160;</td>
			<td>10007-12</td>
			<td>Optimal for dural incision and precision work</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Non-Sterile Scalpel Blades #11</td>
			<td style="width:191px;">FST&#160;</td>
			<td>10011-00</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Surgical Forceps</td>
			<td style="width:191px;">FST&#160;</td>
			<td>11024-18</td>
			<td>The tip of the surgical forceps ensure a firm grip&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Kerrison Bone Punch</td>
			<td style="width:191px;">Aesculap Neurosurgery</td>
			<td style="width:149px;">FF713R</td>
			<td>Must be robust, bite size 3-5 mm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Bone Rongeur</td>
			<td>Aesculap Neurosurgery</td>
			<td style="width:149px;">MD615</td>
			<td>Must be robust, bite size 15 x 5 mm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Bone Rongeur</td>
			<td>Aesculap Neurosurgery</td>
			<td style="width:149px;">FO551R</td>
			<td>Must be robust, bite size 25 x 15 mm&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Bone Chisel</td>
			<td>Lawton</td>
			<td>67-0335</td>
			<td>The size of the chisel head should not exceed 20 mm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Mallet (Hammer)</td>
			<td>Millarco</td>
			<td>5624108</td>
			<td>Weigth 300 g, length 30 cm, head hit area size 2 x 2 cm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Micro-Scissor</td>
			<td>FST&#160;</td>
			<td>14002-14&#160;&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dissector</td>
			<td>Aesculap Neurosurgery</td>
			<td style="width:149px;">OL165R</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">&#160;G&#246;ttingen minipigs</td>
			<td style="width:191px;">&#160;Ellegaard G&#246;ttingen Minipigs A/S, Denmark</td>
			<td style="width:149px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Euthanimal</td>
			<td style="width:191px;"></td>
			<td style="width:149px;"></td>
			<td>pentobarbital</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ketamine</td>
			<td style="width:191px;">Pfizer</td>
			<td style="width:149px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Midazolam&#160;</td>
			<td style="width:191px;">Hameln Pharmaceuticals</td>
			<td style="width:149px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

exposure of the pig cns for histological analysis: a manual for decapitation, skull opening, and brain removal

Pigs have become increasingly popular in large-animal translational neuroscience research as an economically and ethically feasible substitute to non-human primates. The large brain size of the pig allows the use of conventional clinical brain imagers and the direct use and testing of neurosurgical procedures and equipment from the human clinic. Further macroscopic and histological analysis, however, requires postmortem exposure of the pig central nervous system (CNS) and subsequent brain removal. This is not an easy task, as the pig CNS is encapsulated by a thick, bony skull and spinal column. The goal of this paper and instructional video is to describe how to expose and remove the postmortem pig brain and the pituitary gland in an intact state, suitable for subsequent macroscopic and histological analysis.

To prevent the tissue material from drying out, it is recommended to store the removed brain and pituitary in a jar filled with fixative or isotonic saline immediately after macroscopic analysis has been performed. The tissue material may be stored in the fixative for years, whereas storage in isotonic saline, even in a refrigerator, will lead to tissue decay with time.
The removed pituitary may also be directly frozen by immers...

Watch this Scientific Journal Video about Exposure of the Pig CNS for Histological Analysis: A Manual for Decapitation, Skull Opening, and Brain Removal at JoVE.com

Exposure of the Pig CNS for Histological Analysis: A Manual for Decapitation, Skull Opening, and Brain Removal

The goal of this paper and instructional video is to describe how to expose and remove the postmortem pig brain and pituitary gland in an intact state, suitable for subsequent macroscopic and histological analysis.

Pigs have become increasingly popular in large-animal translational neuroscience research as an economically and ethically feasible substitute to ...

In this video, we present how to expose and remove the intact pig brain and the intact pituitary by opening the skull and dura. Ethics statement:Animal anesthesia and euthanasia were performed in accordance with Principles of laboratory animal care"and approved by the Danish Council for Animal Research Ethics. Anesthesia was induced in male Gottingen minipigs by an intramuscular injection of midazolam and ketamine.Five to ten minutes later, when the animals were deeply sedated, an ear vein was cannulated and a lethal overdose of sodium pentobarbital was given intravenously. To secure the animals were completely euthanized, the interdigital pain reflex was tested as previously shown. After proper euthanization, the animals were transcardially perfused with five liters of isotonic saline.All the procedures demonstrated are, accordingly, performed postmortem, excluding the need for welfare precautions necessary for long term anesthesia and post-procedural survival. Instruments used. Heavy scalpel handle number four, and non-sterile scalpel blades, number 23.Good for skin incisions, and soft tissue removal. Scalpel handle number seven, and non-sterile scalpel blades, number 11. Optimal for dural incision, and precision work.Surgical forceps, with a bite that ensures a firm grip. KAIRison Bone Punch. Must be robust, with a bite size of three to five millimeters.Bone rongeur. Must be robust and have a bite size around five to 15 millimeters, for delicate bone work. Larger bone rongeur.With a bite size around 25 times 15 millimeters, for removal of thicker bone pieces. Bone chisel. The head should not be larger than 20 millimeters.A mallet, suitable for the bone chisel. Micro scissors, with a sharp tip.Dissector.Decapitation. The pig is decapitated by making a high circular cervical incision, just below the mandibular angle.Continue the incision, through the soft tissue of the neck. Cut through larynx, and esophagus, until the bone and spinal column is reached. Approximately at the level of the cranio-cervical junction.Advance the incision from the anterior side of the cranio-cervical junction, through the anterior atlantooccipital membrane. Whereby the spinal cord is exposed. Simultaneously, an assistant will pull the pig body away from the pig head, as this will ease the access between the skull base and the first cervical vertebra.Make sure the spinal cord is completely severed, before a forceful extension at the cranio-cervical junction finalizes the decapitation. Skull opening. Place a dorsal longitudinal incision through the skin and underlying soft tissue.From the back of the snout over the vertex of the head and down through the posterior part of the occipital region. Expose the dorsal and posterior part of the skull by removing soft tissue located lateral to the initial incision. Release the temporal muscle from the skull.Clean the posterior occipital bone from soft tissue. Use the posterior interest of the foramen magnum to remove the occipital bone in order to expose the dura-covered cerebellum. Return to the anterior side of the skull and select an entry point in the frontal bone, just in front of the eyes.At this point, use a bone chisel to penetrate the skull, and enter the frontal sinus. Use the extent of the frontal sinus for further removal of the other skull lamina. An inner thin skull lamina covering the cerebrum, is exposed.Open this lamina gently, to expose the dura covering the anterior part of the brain. Continue the bone removal laterally, through the temporal and parietal bone. In order to release the final part of the skull, located between the already-exposed parts of the cerebrum and cerebellum.Use the chisel to break open the remaining posterior skull bone, from one side, just like one opens a door. Brain removal. Lift the dura, for gentle incision close to the sagittal sinus.Use a dural opening for further removal of the dural sac covering the brain. Special care must be taken when removing the dura situated between the cerebrum and the cerebellum. As preservation of this dural leaf will prevent subsequent brain removal.Finally, the dura covering the cerebellum and the lower brain stem is sectioned. Position the pig head vertically, and release the cerebrum by blunt dissection of the olfactory bulbs from the dura-covered skull base. Section the exposed optic chiasm.Follow it by subsequent exposure and section of the pituitary stalk, and the oculomotor nerves. Release the brain stem by sectioning of the lower cranial nerves. Use a chisel to release the brain from the skull base.Pituitary removal. In the skull base, identify the pituitary stalk, with surrounding dural leaf. Cut the dural leaf laterally to the pituitary stalk.Use a dissector to release the pituitary, and lift it out of the pituitary fossa. Results and conclusion. The described technique enables safe removal of the postmortem pig brain and pituitary, in approximately 15 to 20 minutes.The obtained tissue may be submitted for microscopic analysis followed by oriented sectioning, into smaller brain slabs. The slabs are suitable for chemical analysis, or further histological processing.

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