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大鼠脑组织规范性取材专业技术岗位汇报大鼠脑组织规范性取材专业技术岗位汇报秦秋平陆姮磊秦秋平2013-09-25上海药物所药物安全评价研究中心20161012中国科学院上海药物研究所药物安全评价研究中心2016-10-12主要内容主要内容大鼠脑的解剖结构大鼠脑的解剖结构不同的取材方式不同的取材方式常见人工假象常见人工假象不同取材方式之间的比较不同取材方式之间的比较脑的解剖结构脑的解剖结构脑的最前端(嗅球)位于眶间最狭窄处横嵴-嗅球与大脑分界处大脑Bregma前囟点大脑小脑分界处(前缘骨缝)松果体解剖时最容易损伤三处松果体解剖时,最容易损伤三处:嗅球、垂体和小脑绒球脑的解剖结构脑的解剖结构•嗅球•前额皮质前额皮质•尾状核•穹隆-胼胝体后端向下弯纤维束•胼胝体-两大脑半球之间弧形白色神经纤维带•海马•海马•丘脑上部•丘脑下部•横窦•小脑•嗅球脚•嗅球脚•前联合(连接同一半球不同部位)•梨状叶•视神经交叉•乳状体脚间窝•脚间窝•副海马皮质区•锥体重要的神经系统功能结构重要的神系统功能结构Regionsthatperformcriticalneurologicalfunctionsshouldbesampledroutinely.Motor sensoryCognitionHomeostasiscommon neurotoxicitytargetsitesCertainstructuresshouldbesampledregularly:Caudateputamen纹状体Cerebellum小脑Cerebellum小脑Cerebralcortex(frontal额区,parietal顶区,cingulate-扣带回,temporal颞区,occipital枕区,retrospenial压后皮质,piriform梨状叶,entorhinal内皮层)Choroidplexus脉络丛Eye(withopticnerve)眼含视神经Hi海马Hippocampus海马(CornuAmmonis海马角andDentategyrus齿状回)Hypothalamus下丘脑medullaoblongata延髓(Trigeminal三叉神经,Vagal&Facialnuclei迷走和面部神经,AreaPostrema后极区)Midbrain中脑(RostralandCaudalcolliculus前丘和后丘,MedialGeniculatenuclei内侧膝状体核,Rednucleus红核,Raphenucleus中缝核,Substantia中脑(前和后,内侧膝状体核,红核,p中缝核,nigra黑物质)Nerve神经olfactorybulb(rodentsonly)-嗅球Pons脑桥Pons脑桥spinalcord-脊髓Thalamus-丘脑固定和制片固定和制片•解剖时剥离头盖骨固定于10%中性福尔马林溶液24小时后换液•解剖时剥离头盖骨,固定于10%中性福尔马林溶液,24小时后换液•继续固定2-3天后进行取材。•取材后固定2天,进行脱水处理。•常规脱水60%乙醇溶液开始。•HE染色封片•封片1)Cerebrumattheopticchiasm(视交叉神经处大脑))p2)Cerebrumatthebaseoftheposteriorhypothalamus(下丘脑后部处的大脑)3)Mid-cerebellumandmedullaoblongata(中脑和延髓)Optional:4)Ponsatthemiddleofitsprotrusion(脑桥)(recommendedforspecialinvestigationsofthepons.)5)Asampleofthecranialcervicalcordcanberemovedtogetherwiththebrain(location5).Alternatively,itcanbetakentogetherBrain,lateralviewwiththecranialvertebralcolumntoavoidartifacts.1.thefirstsectionatthelevelofopticchiasmincludingthebasalganglia,septum,cortex,anteriorhypothalamus(包括基底核、隔膜、皮质和下丘脑前部)2.thesecondsectionatthelevelofhippocampuscontainingthecortexandbrainstematthetransitionofdiencephalontomesencephalon(间脑到中脑的过渡)Brain,ventralviewmesencephalon(间脑到中脑的过渡)3.thethirdsectioncontainingthecerebellumandbrainstem(medullaoblongata)(延髓).Reference:Revisedguidesfororgansamplingandtrimminginratsandmice.Section1cerebrumSection 1, cerebrumAc:anteriorcommissure(前联合)Cc:cerebralcortex大脑皮质Cca:corpuscallosum(胼胝体)O:opticchiasm(视神经交叉)p(视神经交叉)Sr:striatum(纹状体)CP–CaudatePutamen(尾状核)(basalganglia)SN–SeptalNuclei(隔核)SNCPSPp(隔核)(limbicsystem)SP-septum隔膜HY-hypothalamus下丘脑HYAtthelevelofopticchiasmincludingthebasalganglia,septum,cortex,anteriorhypothalamus(includingthecerebrumatthefrontalcorticallevel)(typicallyrostralforebrain[cerebralcortexandbasalnuclei](typicallyrostralforebrain[cerebralcortexandbasalnuclei],Section2cerebrumSection 2, cerebrumCc:cerebralcortexCc:cerebralcortexCca:corpuscallosumH:hypothalamusHc:hippocampusT:thalamusChPRST:thalamusChP–ChoroidPlexusRS–RetrosplenialCortex(压后皮质)ChPthesecondsectionatthelevelofhippocampuscontainingthecortexandbrainstematthetransitionofdiencephalontomesencephalon(thalamusatthemid-infundibularlevel)Caudalforebrain[cerebralcortexandhippocampuswitheitherdiencephalonorrostralmidbrain]Section3:cerebellumandmedullaoblongataSection 3:cerebellum and medulla oblongataC:cerebellumC:cerebellumMo:medullaoblongataSection4:cerebrumandponsSection 4:cerebrum and ponsCc:cerebralcortexCc:cerebralcortexCca:corpuscallosumHc:hippocampusP:ponsCCAP:ponsT:thalamushindbrain[usuallycerebellumwithponsand/orcerebellumwithmedullaoblongata.hindbrain[usuallycerebellumwithponsand/orcerebellumwithmedullaoblongata.Ponsatthemiddleofitsprotrusion(recommendedforspecialinvestigationsofthepons.)BrainmatirxBrain matirxToachievepreciseverticalcutsbrainmatrixmouldscanbeusedDifferentsizesToachievepreciseverticalcuts,brainmatrixmouldscanbeused.Differentsizesareavailablefordifferentlaboratoryspecies.Forrodents,thebraincanbeslicedat1mmintervals.Reference:RossellaD.Neuroanatomy-basedMatrixguidedTrimmingProtocolfortheRatBrain,ToxicologicPhlXX182014Pathology,XX:1-8,2014.prefrontalcortex(levelA)前额皮质p()前额皮质throughthefrontalpole(levelB)额极attheopticchiasm(levelC)视神经交叉p()视神交叉levelrostraltotheinfundibulum(levelD)漏斗管levelrostraltothecaudalmargin乳头体尾部g乳头体尾部ofthemamillarybody(levelE),throughthemidbrain(3rdcranialnervelevel;levelF)中脑rostraltothemidpointofthecerebellum(levelG),小脑绒球throughthecaudalportionofthecerebellum(levelH).Foreachlevel,anatomicalstructuresofinterestareshown:aca=anteriorcommissure;Acb=accumbensnucleus;Amy=amygdala;Arc=arcuatenuclei;Au=auditorycortex;B=basalnucleus;CA1CA2CA3=cornuammonis12and3;cc=corpusAmy=amygdala;Arc=arcuatenuclei;Au=auditorycortex;B=basalnucleus;CA1,CA2,CA3=cornuammonis1,2and3;cc=corpuscallosum;Cer=cerebellum;Cg=cingulatecortex;cg=cingulum;cic=inferiorcolliculicommissure;cp=cerebralpeduncles;Cpu=caudateputamen;csc=superiorcolliculicommissure;DG=dentategyrus;DR=dorsalraphenucleus;Ect=ectorhinalcortex;Ent=entorhinalcortex;Fl=flocculus;fmi=forcepsminorofthecorpuscallosum;f=fornix;fi=fimbria;GP=globuspallidus;Hb=habenularnuclei;Hyp=hypothalamus;ic=internalcapsule;IC=inferiorcolliculi;ICj=Callejaislands;icp=infer
本文标题:04.陆�磊--大鼠脑组织规范性取材
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