저자는 감사로 부인 트린 W 미켈슨의 숙련 된 지원, 피팅 부인 리자 M. 및 Påskehøjgaard에서 직원을 인정합니다. 덴마크 의학 연구위원회 (Medical Research Council)의 룬드 벡 재단, 그리고 노보 노 디스크 재단은 재정적으로 연구를 지원했다.

동물 마취 및 euthanesia은 (NIH 간행물 번호 86-23, 1985 개정) 동물 연구 윤리에 대한 덴마크위원회의 승인을 &quot;실험 동물 관리의 원칙&quot;에 따라 수행되었다. 1. 악기 <ol><li> 악기 비디오에서 제시 및 재료의 표에 열거를 수집합니다. </li></ol> 2. 목 베기 주 : 마취 미다 졸람 5 ㎖ (5 ㎎ / ㎖) 및 5 mL의 케타민 (25 ㎎ / ㎖)의 근육 내 주사에 의해 유도 하였다. 동물을 깊게 마취 때 50-10 분 후에, 귀 정맥 삽관시키고, 나트륨 펜토 바르 비탈의 과량 치사 (체중 100 밀리그램 / kg)을 (200 ㎎ / ㎖)을 정맥 받았다. Ettrup 등에 의해 도시 된 바와 같이, 동물은 안락사 완전히 것을 보장하기 위해, 인터 통증 플렉스 시험 하였다. (2011) 20. 에 설명 된대로 전체 euthanization 확보했다Ettrup 등에 의해 입증 된 바와 같이하고, 등장 식염수 5 L와 transcardial 관류 하였다 위에 윤리 문. (2011) 20. 모든 입증 절차는 장기 마취 및 postprocedural 생존에 필요한 복지주의가 필요하지 않습니다, 사후을 수행한다. <ol><li> 다만 하악 각도 (도 1a) 아래 수술 메스를 사용하여, 높은 원형 자궁 절개하여 돼지 참살. </li><li> 뼈 척추가 약 craniocervical 접합의 수준에 도달 할 때까지 아직도 수술 메스, 후두 및 식도를 포함한 목의 부드러운 조직을 통해 전방 절개를 계속합니다. </li><li> (아틀라스 전치부 호 위의 craniocervical 접합의 전방 측으로부터 외과 용 메스로 절단을 향상하고, 전방 atlantooccipital 막을 통해하여 척추관 척수 노출그림 1B). 동시에 두개골베이스와 첫 번째 경추 사이의 접근을 용이하게하기 위해 돼지 머리에서 멀리 돼지 몸을 끌어 보조를 부탁드립니다. </li><li> 경막 주머니와 척수 (그림 1B)를 통해 수술 절개를 계속합니다. 척수의 전체 횡단면이 달성 될 수 있도록 특별한주의하십시오. 참고 : 잘린 프로세스의 다음 단계 동안 척수와 뇌에 불필요한 마찰이 발생할 수 있습니다 이전 단계를 수행하지 않으면. </li><li> 강제 섹션 레벨 (도 1C)에 접합 cranocervical 연장. 동시에, 후두부 관절 구 및 아틀라스의 상부 명료 프로세스 사이의 관절을 해제 섹션에 나머지 atlantooccipital 인대 외과 용 메스를 사용한다. 몸에서 돼지 머리를 분리합니다. </li></ol><img alt="그림 1" src="/files/ftp_upload/55511/55511fig1.jpg" style="font-size: 14px;" /> 그림 1 : 미니 피그의 참수. (A) 목 절개 (화살표 하악 각도). (; OC, 후두부 외과 C1, 아틀라스 전치부 호)에 atlantooccipital 인대 및 craniocervical 접합부 경질 둘러싸인 척수 (SC) 내지 (B) 절개. (C)가 atlantooccipital 관절의 후방 부분은 부 수준에서 강력한 확장 (화살표)에 의해 방출된다. <a href="http://ecsource.jove.com/files/ftp_upload/55511/55511fig1large.jpg" style="font-size: 14px;" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> 3. 해골 열기 <ol><li> 테이블에 돼지 머리를 놓습니다. </li><li> 의 후방 부분을 통해 피부와 머리의 정점에 걸쳐 주둥이의 뒷면에서 기본 부드러운 조직을 통해 수술 메스와 지느러미 세로 절개를, 아래로후두부. </li><li> 외과 용 메스 절개 초기에 측 방향 위치 연조직을 제거하여 두개골의 등쪽과 후방 부분을 노출시킨다. </li><li> 외과 용 메스와 두개골 (도 2A)로부터 임시 좌우 근육을 해제. 후방 뒤통수 뼈는 연부 조직의 청소되어 있는지 확인합니다. </li><li> Kerrison 뼈와 뼈 rongeurs 펀치와 후두부 뼈를 분리하고 경질 피복 소뇌 (도 2b)를 노출시키는 난원 매그넘의 후방 입구를 사용한다. </li><li> 바로 눈 앞에서, 두개골의 노출 전방 측에 돌아가서 정면 뼈의 엔트리 포인트를 선택합니다. 이 시점에서, 두개골을 관통하고 전두엽 부비동 (그림 2C)을 입력 망치로 뼈 조각도를 사용합니다. </li><li> 뼈 또는 뼈 rongeur 펀치 외 두개골 박판 dorsoposterior의 제거를 촉진 할 전두동의 범위를 사용하고, 내부 얇은 노출대뇌 (도 2D)를 덮는 박판 두개골 뼈. </li><li> 부드럽게 경질 피복 뇌 (도 2E)을 노출시키는 해머와 뼈 조각도와 전방 내부 뼈 두개골 박판을 연다. </li><li> 횡 경질 피복 대뇌 및 소뇌 (도 2F)에 이미 노출 된 부분 사이에 두개골의 최종 dorsoposterior 부분을 분리하기 위해 시간적 마루 뼈 통해 뼈 정 및 뼈 rongeur을 사용하여 골 삭제를 계속한다. 참고 :이 절차의 마지막 단계에서, 가능한 한 자주 하나가 문을 열어처럼 한쪽에서 열린 남아있는 후방 두개골 뼈를 파괴하기 위해 끌을 사용합니다. </li></ol><img alt="그림 2" src="/files/ftp_upload/55511/55511fig2.jpg" style="opacity: 0.9; font-size: 14px;" /> 도 2 : 미니 피그 두개골 개구. (A)가 노출되는 dorsoposterio후두 시간적 근육의 제거를 포함하여 R 두개골 표면. 후두골의 (B)를 제거 (CB, 경질 피복 소뇌, OB, 후두골, OC, 후두부 외과 및 SC 척수). (C) 망치 뼈 치즐 전방 두개골을 관통하고 눈의 수준에서 전두동를 입력하기 위해 사용된다. (D) 전방 공동 (FS)의 정도는 내부 얇은 뼈 박판 (2) 뇌를 덮는 노출 (1) 외측 두꺼운 두개골 뼈를 제거하기 위해 사용된다. 경질 피복 대뇌 (화살표)을 노출 얇은 뼈의 박판 (E) 제거. (F)는 마지막으로, 망치와 뼈 시추 좌우 전방 및 후방 두개골 개구를 연결하는 데 사용된다. <a href="http://ecsource.jove.com/files/ftp_upload/55511/55511fig2large.jpg" style="font-size: 14px;" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> 4. 뇌 Remov알 <ol><li> 경질을 들어 올려 미세 수술 메스 (그림 3A)를 사용하여 정맥 우수한 시상 동에 부드러운 절개 가까이를 만드는 수술 집게를 사용합니다. </li><li> 상기 뇌의 등쪽 표면을 덮는 경질를 열고 마이크로 가위 또는 경질 칼을 사용한다. 참고 : 소뇌 tentorium (그림 3B)에 해당하는 경질을 제거 할 때 특별한주의가 이후의 뇌 제거를 방지 할이 경막 잎의 보존으로,주의해야합니다. </li><li> 돼지 머리를 수직으로 (그림 3C)를 배치합니다. </li><li> 뇌 캐비티 (도 3d)의 경질 피복 층에서 후각 망울의 무딘 절개하여 대뇌 ventroanterior를 해제 뼈 정 또는 해부학자 사용. </li><li> 섹션에 노출 광학 chiasm (그림 3E)을 미세 수술 메스를 사용합니다. 뇌하수체 줄기와 안구 운동 신경을 노출하고 섹션을 참조하십시오. </li><li> 소호을 절편하여 복부 뇌간을 해제미세 수술 메스와 WER 뇌신경 (그림 3 층). 경막 소뇌 tentorium 완전히 (그림 3B) 절개 것을이 경막 잎 그렇지 않으면 해제 과정에서 뇌간을 통해 절단하므로, 확인합니다. </li></ol><img alt="그림 3" src="/files/ftp_upload/55511/55511fig3.jpg" style="font-size: 14px;" /> 그림 3 : 미니 피그 뇌 제거. 수술 집게와 경막 칼 (A) 경막 개방. (B) 관리는 완전히 대뇌 및 소뇌 사이에 경막 리프 (화살표), 절개주의해야한다. (C) 돼지 머리 두개골베이스 구조 나은 시각화 중력 뇌의 의도 용량을 지원하기 위해서는 수직으로 배치된다. (D)는 해부학자 또는 뼈 속이다은 경질 커버 ③에서 평활 수단에 의해 후각 망울을 완화하는 데 사용D 두개골베이스. (E)을 박리는 광학 chiasm (화살표), 줄기 누 두부 및 안구 운동 신경의 노출 단면에 대한 두개골베이스를 따라 후방 방향으로 계속된다. 그들은 뇌간의 복부 표면으로부터 출발로 (F) 뇌 릴리스 하부 뇌신경의 단면 완성된다 (, IV, 활차 신경, III, 눈 돌림 신경 V, 삼차 신경 및 VI를 외전 신경). <a href="http://ecsource.jove.com/files/ftp_upload/55511/55511fig3large.jpg" style="font-size: 14px;" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> 5. 뇌하수체 제거 <ol><li> 뇌하수체 줄기 두개골 층 (도 4a)의 주변 경막 잎합니다 (diapragma의 sellae)를 식별. </li><li> 미세 수술 용 메스 (도 4b)을 이용하여 뇌하수체 스토킹 경막 리프 횡 절개. </li><li> 구덩이를 해제하는 해부학자를 사용하여uitary 및 뇌하수체 포사 (그림 4C)에서 들어 올립니다. </li></ol><img alt="그림 4" src="/files/ftp_upload/55511/55511fig4.jpg" style="opacity: 0.9; font-size: 14px;" /> 그림 4 : 미니 피그의 뇌하수체 제거. (A) 뇌하수체 포사 (*)는 두개골 바닥에서 식별된다 (1, 후각 망울 2, 광학 chiasm 및 PF, 사후 뇌 포사 참조). (B) 경막 커버 (sellar diagphragm (화살표)) 측 방향으로 절개되어있다. (C)이 뇌하수체 (화살표)는 해부학자로 방출되어 뇌하수체 포사 밖으로 들어 올려진다. 스케일 바 (AC)는 10mm를 =. <a href="http://ecsource.jove.com/files/ftp_upload/55511/55511fig4large.jpg" style="font-size: 14px;" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a>

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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. R., Gjedde, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kinetics+of+the+uptake+and+distribution+of+the+dopamine+D2/3+agonist+(R)-N-[1-11C]n-propylnorapomorphine+in+brain+of+healthy+and+MPTP-poisoned+Gottingen+miniature+pigs.">Kinetics of the uptake and distribution of the dopamine D2/3 agonist (R)-N-[1-11C]n-propylnorapomorphine in brain of healthy and MPTP-poisoned Gottingen miniature pigs.</a> Nucl Med Biol. 30 (5), 547-553 (2003).</li><li>Jensen, K. N., Deding, D., 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=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. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+strategies+for+embedding,+orientation+and+sectioning+of+small+brain+specimens+enable+direct+correlation+to+MR-images,+brain+atlases,+or+use+of+unbiased+stereology.">New strategies for embedding, orientation and sectioning of small brain specimens enable direct correlation to MR-images, brain atlases, or use of unbiased stereology.</a> J Neurosci Methods. 108, 153-159 (2001).</li><li>Bjarkam, C. R., S&#248;rensen, J. C., 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=Distribution+and+morphology+of+serotonin-immunoreactive+axons+in+the+hippocampal+region+of+the+New+Zealand+white+rabbit.+I.+Area+dentata+and+hippocampus+proper.">Distribution and morphology of serotonin-immunoreactive axons in the hippocampal region of the New Zealand white rabbit. I. Area dentata and hippocampus proper.</a> Hippocampus. 13 (1), 21-37 (2003).</li><li>Bjarkam, C. R., Glud, A. N., Orlowski, D., S&#248;rensen, J. C., Palomero-Gallagher, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+telencephalon+of+the+minipig,+cytoarchitecture+and+cortical+surface+anatomy.">The telencephalon of the minipig, cytoarchitecture and cortical surface anatomy.</a> Brain Struct Funct. , (2016).</li><li>Boltze, J., Nitzsche, B., Geiger, K. D., Schoon, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Histopathological+investigation+of+different+MCAO+modalities+and+impact+of+autologous+bone+marrow+mononuclear+cell+administration+in+an+ovine+stroke+model.">Histopathological investigation of different MCAO modalities and impact of autologous bone marrow mononuclear cell administration in an ovine stroke model.</a> Transl Stroke Res. 2, 279-293 (2011).</li><li>Jortner, B. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+return+of+the+dark+neuron.+A+histological+artifact+complicating+contemporary+neurotoxicologic+evaluation.">The return of the dark neuron. A histological artifact complicating contemporary neurotoxicologic evaluation.</a> Neurotoxicology. 27, 628-634 (2006).</li></ol>

저자가 공개하는 게 없다.

대부분의 실험 연구는 신경 같은 CNS에 대한 액세스가 얇은 skull- 및 경막의 두께에 의해 용이하게 마우스 및 래트와 같은 작은 동물 종에서 수행된다. 그러나, 돼지 1, 4, 8, 양 (32) 및 비 - 인간 영장류와 같은 큰 실험 동물에서 이러한 구조의 상당한 두께 두개골 뼈 제거를위한 강력한 도구 (재료의 테이블) 적절...

<html> 돼지 번역 상 신경 과학 연구는 지난 20 년 동안 점점 더 인기를 끌고있다. 돼지 뇌의 크기가 커서 기존의 임상 뇌 이미 저들과 직접 사용하여 인간 임상 1, 2, 3, 4, 5, 6, 7, 8에서 신경 외과 절차 및 장비 테스트의 사용을 가능하게한다. 지난 20 년 동안, 돼지, 특히 minipigs (예를 들어, 괴팅겐 미니 피그) 등 줄기 세포 이식과 같은 neuromodulatory 치료 방법을 검사하는 데 사용되었다; 바이러스 벡터 형질; 파킨슨 질환, 비만, 우울증, 알츠하이머 병 2, 6 향하는 깊은 뇌 자극,= &quot;외부 참조&quot;&gt; 9, 10, 11, 12, 13, 14, 15, 16, 17. 이는 미니 피그 CNS 3, 18, 19, 20, 21을 조작 정위 수술 방법을 개발 하였다되었다. 제정 CNS의 변경 (PET 10, 13, 22, 24, 23 MR), 방광 내압 11, 12, 25 뇌 영상을 이용하여 살아있는 동물에서 평가되어, 보행 분석17, 신경 평가 9, 17, 및 조직학 분석 stereological 14, 15, 17, 26, 27, 31에 기초하여 사후 검사. 그러나 사후 분석은 두께, 뼈 두개골과 돼지 뇌를 둘러싸고 취재 섬유 경막로 노출 쉬운 일이 아니다 돼지 두뇌의 제거가 필요합니다. 이 종이와 교육 비디오의 목표는 사후 돼지 뇌와 뇌하수체 비 전동 수술 도구를 사용하여 15 ~ 20 분에 온전한 상태로 노출을 제거 할 수있는 방법을 설명하는 것입니다. 미니 피그 뇌하수체에 대한 해부학 적 연구에 사용 된 교육 비디오 및 사진 그림은 남성 minipigs (20-25kg : : 6 개월, 체중 세)를 보여줍니다.

<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

돼지는 인간이 아닌 영장류에 대한 경제적, 윤리적으로 가능한 대안으로 대형 동물 번역의 신경 과학 연구에 점점 더 인기를 끌고있다. 돼지의 큰 뇌의 크기는 기존의 임상 뇌 이미 저와 인간의 병원에서 직접 사용 및 신경 외과 절차 및 장비의 테스트를 사용할 수 있습니다. 또한, 육안 및 조직 학적 분석은, 그러나, 돼지 중추 신경계 (CNS) 및 후속 뇌 제거 사후 노광을 필요로한다. 이것은 CNS는 두께, 뼈 두개골과 척추에 의해 캡슐화 된 돼지로 쉬운 일이 아니다. 이 종이와 교육 비디오의 목표는 노출과 사후 돼지 뇌와 이후의 거시 조직 학적 분석에 적합한 손상 상태에서 뇌하수체를 제거하는 방법을 설명하는 것입니다.

마르지 티슈 재료를 방지하기 위해,이 고정 제 또는 거시적 분석이 수행 된 직후에 등장 성 식염수로 채워진 병의 뇌 및 뇌하수체 제거를 저장하도록 권고한다. 조직 재료는, 등장 성 식염수에 보관 반면 년 동안 정착에 저장 될 수있다, 심지어 냉장고에, 시간과 함께 조직의 붕괴로 이어질 것입니다. 본래 돼지 뇌 직접 동결 <s...

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

이 종이와 교육 비디오의 목표는 노출 이후 거시 및 조직 학적 분석에 적합한 손상 상태에 돼지 뇌와 뇌하수체 부검을 제거하는 방법을 설명하는 것입니다.

조직 학적 분석을위한 돼지 CNS의 노출 : 목 베기, 두개골 열기위한 매뉴얼 및 뇌 제거

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

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The Axial Skeleton

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15.3K 조회수.  Aalborg University Hospital.  이 종이와 교육 비디오의 목표는 노출 이후 거시 및 조직 학적 분석에 적합한 손상 상태에 돼지 뇌와 뇌하수체 부검을 제거하는 방법을 설명하는 것입니다. 

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

A Polished and Reinforced Thinned-skull Window for Long-term Imaging of the Mouse Brain

In vivo imaging of cortical function requires optical access to the brain without disruption of the intracranial environment. We present a method to form a polished and reinforced thinned skull (PoRTS) window in the mouse skull that spans several millimeters in diameter and is stable for months. The skull is thinned to 10 to 15 &mu;m in thickness with a hand held drill to achieve optical clarity, and is then overlaid with cyanoacrylate glue and a cover glass to: 1) provide rigidity, 2) inhibit bone regrowth and 3) reduce light scattering from irregularities on the bone surface. Since the skull is not breached, any inflammation that could affect the process being studied is greatly reduced. Imaging depths of up to 250 &mu;m below the cortical surface can be achieved using two-photon laser scanning microscopy. This window is well suited to study cerebral blood flow and cellular function in both anesthetized and awake preparations. It further offers the opportunity to manipulate cell activity using optogenetics or to disrupt blood flow in targeted vessels by irradiation of circulating photosensitizers.

We present a method to form an imaging window in the mouse skull that spans millimeters and is stable for months without inflammation of the brain. This method is well suited for longitudinal studies of blood flow, cellular dynamics, and cell/vascular structure using two-photon microscopy.

마우스 뇌의 장기 이미징을위한 세련된와 철근 Thinned-해골 창

In vivo imaging of cortical function requires optical access to the brain without disruption of the intracranial environment. We present a method to form ...

연구

신경과학

Co-analysis of Brain Structure and Function using fMRI and Diffusion-weighted Imaging

The study of complex computational systems is facilitated by network maps, such as circuit diagrams. Such mapping is particularly informative when studying the brain, as the functional role that a brain area fulfills may be largely defined by its connections to other brain areas. In this report, we describe a novel, non-invasive approach for relating brain structure and function using magnetic resonance imaging (MRI). This approach, a combination of structural imaging of long-range fiber connections and functional imaging data, is illustrated in two distinct cognitive domains, visual attention and face perception. Structural imaging is performed with diffusion-weighted imaging (DWI) and fiber tractography, which track the diffusion of water molecules along white-matter fiber tracts in the brain (Figure 1). By visualizing these fiber tracts, we are able to investigate the long-range connective architecture of the brain. The results compare favorably with one of the most widely-used techniques in DWI, diffusion tensor imaging (DTI). DTI is unable to resolve complex configurations of fiber tracts, limiting its utility for constructing detailed, anatomically-informed models of brain function. In contrast, our analyses reproduce known neuroanatomy with precision and accuracy. This advantage is partly due to data acquisition procedures: while many DTI protocols measure diffusion in a small number of directions (e.g., 6 or 12), we employ a diffusion spectrum imaging (DSI)1, 2 protocol which assesses diffusion in 257 directions and at a range of magnetic gradient strengths. Moreover, DSI data allow us to use more sophisticated methods for reconstructing acquired data. In two experiments (visual attention and face perception), tractography reveals that co-active areas of the human brain are anatomically connected, supporting extant hypotheses that they form functional networks. DWI allows us to create a "circuit diagram" and reproduce it on an individual-subject basis, for the purpose of monitoring task-relevant brain activity in networks of interest.

We describe a novel approach for simultaneous analysis of brain function and structure using magnetic resonance imaging (MRI). We assess brain structure with high-resolution diffusion-weighted imaging and white-matter fiber tractography. Unlike standard structural MRI, these techniques allow us to directly relate anatomical connectivity to functional properties of brain networks.

fMRI 및 확산 - 가중 영상을 사용하여 뇌의 구조와 기능의 공동 분석

The study of complex computational systems is facilitated by network maps, such as circuit diagrams. Such mapping is particularly informative when ...

A Comprehensive Protocol for Manual Segmentation of the Medial Temporal Lobe Structures

The present paper describes a comprehensive protocol for manual tracing of the set of brain regions comprising the medial temporal lobe (MTL): amygdala, hippocampus, and the associated parahippocampal regions (perirhinal, entorhinal,&#160;and parahippocampal proper). Unlike most other tracing protocols available, typically focusing on certain MTL areas (e.g., amygdala and/or hippocampus), the integrative perspective adopted by the present tracing guidelines allows for clear localization of all MTL subregions. By integrating information from a variety of sources, including extant tracing protocols separately targeting various MTL structures, histological reports, and brain atlases, and with the complement of illustrative visual materials, the present protocol provides an accurate, intuitive, and convenient guide for understanding the MTL anatomy. The need for such tracing guidelines is also emphasized by illustrating possible differences between automatic and manual segmentation protocols. This knowledge can be applied toward research involving not only structural MRI investigations but also structural-functional colocalization and fMRI signal extraction from anatomically defined ROIs, in healthy and clinical groups alike.

The present work provides a comprehensive set of guidelines for manually tracing the medial temporal lobe (MTL) structures. This protocol can be applied to research involving structural and/or combined structural-functional magnetic resonance imaging (MRI) investigations of the MTL, in both healthy and clinical groups.

내측 측두엽 구조의 수동 분할을위한 포괄적 인 프로토콜

The present paper describes a comprehensive protocol for manual tracing of the set of brain regions comprising the medial temporal lobe (MTL): amygdala, ...

Analyzing Synaptic Modulation of Drosophila melanogaster Photoreceptors after Exposure to Prolonged Light

The nervous system has the remarkable ability to adapt and respond to various stimuli. This neural adjustment is largely achieved through plasticity at the synaptic level. The Active Zone (AZ) is the region at the presynaptic membrane that mediates neurotransmitter release and is composed of a dense collection of scaffold proteins. AZs of Drosophila melanogaster (Drosophila) photoreceptors undergo molecular remodeling after prolonged exposure to natural ambient light. Thus the level of neuronal activity can rearrange the molecular composition of the AZ and contribute to the regulation of the functional output.
Starting from the light exposure set-up preparation to the immunohistochemistry, this protocol details how to quantify the number, the spatial distribution, and the delocalization level of synaptic molecules at AZs in Drosophila photoreceptors. Using image analysis software, clusters of the GFP-fused AZ component Bruchpilot were identified for each R8 photoreceptor (R8) axon terminal. Detected Bruchpilot spots were automatically assigned to individual R8 axons. To calculate the distribution of spot frequency along the axon, we implemented a customized software plugin. Each axon&#39;s start-point and end-point were manually defined and the position of each Bruchpilot spot was projected onto the connecting line between start and end-point. Besides the number of Bruchpilot clusters, we also quantified the delocalization level of Bruchpilot-GFP within the clusters. These measurements reflect in detail the spatially resolved synaptic dynamics in a single neuron under different environmental conditions to stimuli.

Analyzing Synaptic Modulation of Drosophila melanogaster Photoreceptors after Exposure to Prolonged Light

Here we show how to quantify the number and spatial distribution of synaptic active zones in Drosophila melanogaster&#160;photoreceptors, highlighted with a genetically encoded molecular marker, and their modulation after prolonged exposure to light.

의 시냅틱 변조 분석<em&gt; 노랑 초파리</em&gt; 장시간 빛에 노출 후 광수

The nervous system has the remarkable ability to adapt and respond to various stimuli. This neural adjustment is largely achieved through plasticity at ...

High-throughput Analysis of Locomotor Behavior in the Drosophila Island Assay

Advances in next-generation sequencing technologies contribute to the identification of (candidate) disease genes for movement disorders and other neurological diseases at an increasing speed. However, little is known about the molecular mechanisms that underlie these disorders. The genetic, molecular, and behavioral toolbox of Drosophila melanogaster makes this model organism particularly useful to characterize new disease genes and mechanisms in a high-throughput manner. Nevertheless, high-throughput screens require efficient and reliable assays that, ideally, are cost-effective and allow for the automatized quantification of traits relevant to these disorders. The island assay is a cost-effective and easily set-up method to evaluate Drosophila locomotor behavior. In this assay, flies are thrown onto a platform from a fixed height. This induces an innate motor response that enables the flies to escape from the platform within seconds. At present, quantitative analyses of filmed island assays are done manually, which is a laborious undertaking, particularly when performing large screens.
This manuscript describes the &#34;Drosophila Island Assay&#34; and &#34;Island Assay Analysis&#34; algorithms for&#160;high-throughput, automated data processing and quantification of island assay data. In the setup, a simple webcam connected to a laptop collects an image series of the platform while the assay is performed. The &#34;Drosophila Island Assay&#34; algorithm developed for the open-source software Fiji processes these image series and quantifies, for each experimental condition, the number of flies on the platform over time. The &#34;Island Assay Analysis&#34; script, compatible with the free software R, was developed to automatically process the obtained data and to calculate whether treatments/genotypes are statistically different. This greatly improves the efficiency of the island assay and makes it a powerful readout for basic locomotion and flight behavior. It can thus be applied to large screens investigating fly locomotor ability, Drosophila models of movement disorders, and drug efficacy.

High-throughput Analysis of Locomotor Behavior in the Drosophila Island Assay

The island assay is a relatively new, cost-effective assay that can be used to evaluate the basic locomotor behavior of Drosophila melanogaster. This manuscript describes algorithms for automatic data processing and objective quantification of island assay data, making this assay a sensitive, high-throughput readout for large genetic or pharmacological screens.

Drosophila 섬 분석 결과에서 운전 행동의 높은 처리량 분석

Advances in next-generation sequencing technologies contribute to the identification of (candidate) disease genes for movement disorders and other ...

Recording Synaptic Plasticity in Acute Hippocampal Slices Maintained in a Small-volume Recycling-, Perfusion-, and Submersion-type Chamber System

Even though experiments on brain slices have been in use since 1951, problems remain that reduce the probability of achieving a stable and successful analysis of synaptic transmission modulation when performing field potential or intracellular recordings. This manuscript describes methodological aspects that might be helpful in improving experimental conditions for the maintenance of acute brain slices and for recording field excitatory postsynaptic potentials in a commercially available submersion chamber with an outflow-carbogenation unit. The outflow-carbogenation helps to stabilize the oxygen level in experiments that rely on the recycling of a small buffer reservoir to enhance the cost-efficiency of drug experiments. In addition, the manuscript presents representative experiments that examine the effects of different carbogenation modes and stimulation paradigms on the activity-dependent synaptic plasticity of synaptic transmission.

This protocol describes the stabilization of the oxygen level in a small volume of recycled buffer and methodological aspects of recording activity-dependent synaptic plasticity in submerged acute hippocampal slices.

급성 Hippocampal 조각에 작은 볼륨 재활용-, 관류-침수 형 챔버 시스템 유지에 시 냅 스가 소성을 기록

Even though experiments on brain slices have been in use since 1951, problems remain that reduce the probability of achieving a stable and successful ...

Novel Passive Clearing Methods for the Rapid Production of Optical Transparency in Whole CNS Tissue

Since the development of CLARITY, a bioelectrochemical clearing technique that allows for three-dimensional phenotype mapping within transparent tissues, a multitude of novel clearing methodologies including CUBIC (clear, unobstructed brain imaging cocktails and computational analysis), SWITCH (system-wide control of interaction time and kinetics of chemicals), MAP (magnified analysis of the proteome), and PACT (passive clarity technique), have been established to further expand the existing toolkit for the microscopic analysis of biological tissues. The present study aims to improve upon and optimize the original PACT procedure for an array of intact rodent tissues, including the whole central nervous system (CNS), kidneys, spleen, and whole mouse embryos. Termed psPACT (process-separate PACT) and mPACT (modified PACT), these novel techniques provide highly efficacious means of mapping cell circuitry and visualizing subcellular structures in intact normal and pathological tissues. In the following protocol, we provide a detailed, step-by-step outline on how to achieve maximal tissue clearance with minimal invasion of their structural integrity via psPACT and mPACT.

Here, we present two novel methodologies, psPACT and mPACT, for achieving maximal optical transparency and subsequent microscopic analysis of tissue vasculature in the intact rodent whole CNS.

소설 수동 전체 CNS 조직에 광학 투명도의 급속 한 생산을 위한 방법 삭제

Since the development of CLARITY, a bioelectrochemical clearing technique that allows for three-dimensional phenotype mapping within transparent tissues, ...

Chronic Implantation of Whole-cortical Electrocorticographic Array in the Common Marmoset

Electrocorticography (ECoG) allows the monitoring of electrical field potentials from the cerebral cortex with high spatiotemporal resolution. Recent development of thin, flexible ECoG electrodes has enabled conduction of stable recordings of large-scale cortical activity. We have developed a whole-cortical ECoG array for the common marmoset. The array continuously covers almost the entire lateral surface of cortical hemisphere, from the occipital pole to the temporal and frontal poles, and it captures whole-cortical neural activity in one shot. This protocol describes a chronic implantation procedure of the array in the epidural space of the marmoset brain. Marmosets have two advantages regarding ECoG recordings, one being the homologous organization of anatomical structures in humans and macaques, including frontal, parietal, and temporal complexes. The other advantage is that the marmoset brain is lissencephalic and contains a large number of complexes, which are more difficult to access in macaques with ECoG, that are exposed to the brain surface.These features allow direct access to most cortical areas beneath the surface of the brain. This system provides an opportunity to investigate global cortical information processing with high resolutions at a sub-millisecond order in time and millimeter order in space.

We have developed a whole-cortical electrocorticographic array for the common marmoset that continuously covers almost the entire lateral surface of cortex, from the occipital pole to the temporal and frontal poles. This protocol describes a chronic implantation procedure of the array in the epidural space of the marmoset brain.

일반적인 마모 셋에 전체 피 질 Electrocorticographic 배열의 만성 이식

Electrocorticography (ECoG) allows the monitoring of electrical field potentials from the cerebral cortex with high spatiotemporal resolution. Recent ...

Extraction and Dissection of the Domesticated Pig Brain

Use of the pig as a preclinical and translatable animal model has been well-documented and accepted by research fields investigating cardiovascular systems, gastrointestinal systems, and nutrition, and the pig is increasingly being used as a large animal model in neuroscience. Furthermore, the pig is an accepted model to study neurodevelopment as it displays brain growth and development patterns similar to what occurs in humans. As a less common animal model in neuroscience, surgical and dissection procedures on pigs may not be as familiar or well-practiced among researchers. Therefore, a standardized visual protocol detailing consistent extraction and dissection methods may prove valuable for researchers working with the pig. The following video showcases a technique to remove the pig brain while keeping the cortex and brainstem intact and reviews methods to dissect several commonly investigated brain regions including the brainstem, cerebellum, midbrain, hippocampus, striatum, thalamus, and medial prefrontal cortex. The purpose of this video is to provide researchers with the tools and knowledge necessary to consistently perform a brain extraction and dissection on the four-week-old pig.

This protocol details the technique for removal of the pig brain in its entirety and dissection of several brain regions commonly studied in neuroscience.

길들여진 돼지 뇌의 추출 및 해부

Use of the pig as a preclinical and translatable animal model has been well-documented and accepted by research fields investigating cardiovascular ...

In vivo Imaging of Fully Active Brain Tissue in Awake Zebrafish Larvae and Juveniles by Skull and Skin Removal

Understanding the ephemeral changes that occur during brain development and maturation requires detailed high-resolution imaging in space and time at cellular and subcellular resolution. Advances in molecular and imaging technologies have allowed us to gain numerous detailed insights into cellular and molecular mechanisms of brain development in the transparent zebrafish embryo. Recently, processes of refinement of neuronal connectivity that occur at later larval stages several weeks after fertilization, which are for example control of social behavior, decision making or motivation-driven behavior, have moved into focus of research. At these stages, pigmentation of the zebrafish skin interferes with light penetration into brain tissue, and solutions for embryonic stages, e.g., pharmacological inhibition of pigmentation, are not feasible anymore.
Therefore, a minimally invasive surgical solution for microscopy access to the brain of awake zebrafish is provided that is derived from electrophysiological&#160;approaches. In teleosts, skin and soft skull cartilage can be carefully removed by micro-peeling these layers, exposing underlying neurons and axonal tracts without damage. This allows for recording neuronal morphology, including synaptic structures and their molecular contents, and the observation of physiological changes such as Ca2+ transients or intracellular transport events. In addition, interrogation of these processes by means of pharmacological inhibition or optogenetic manipulation is feasible. This brain exposure approach provides information about structural and physiological changes in neurons as well as the correlation and interdependence of these events in live brain tissue in the range of minutes or hours. The technique is suitable for in vivo brain imaging of zebrafish larvae up to 30 days post fertilization, the latest developmental stage tested so far. It, thus, provides access to such important questions as synaptic refinement and scaling, axonal and dendritic transport, synaptic targeting of cytoskeletal cargo or local activity-dependent expression. Therefore, a broad use for this mounting and imaging approach can be anticipated.

Here we present a method to image the zebrafish embryonic brain in vivo upto larval and juvenile stages. This microinvasive procedure, adapted from electrophysiological approaches, provides access to cellular and subcellular details of mature neuron and can be combined with optogenetics and neuropharmacological studies for characterizing brain function and drug intervention.

두개골과 피부 제거에 의해 깨어 얼룩말 물고기 애벌레와 청소년에서 완전히 활성 뇌 조직의 생체 내 이미징

Understanding the ephemeral changes that occur during brain development and maturation requires detailed high-resolution imaging in space and time at ...