样品制备科大组蛋白质科学研究平台生物成像中心

电镜成像申请请发送 扫描电镜成像请发送 jianguozha样品申请请发送 样品：申请书登记接收—审核结果—通知送样—用户送样—制样完不超过第二个自然周的机时。同样，区域中心内课题组样品优先级大样品技术服务时间承诺：对区域中心内用户，从实际接收合格样品到定，报签字确认。用户在生物成像中心完成科研工作、获取研究数据并据此相关成（、著作等，应在成果中对生物成像中心予以致谢（详细规定参见生的信息，仪器介绍和样品信息等，生物成像中心常规样品电镜成 免疫标记电镜成低温电镜成 扫描电镜成荧光显微 超分辨荧光显微镜图像处 其课题背CentleinCrispr-cas9CentleinhTERT-RPE1本课题组希望通过透射电镜技术，观察野生型和Centlein敲除细胞中原进一步的与纤关疾病研究有重要意义。泡）希望达到的效果如下图例1中所示RepresentativeelectronmicrographsofRPEcellM-centrioles.Scalebar,200nm.参考方法：ElectronmicroscopyprocessinginRPEcells.Briefly,cellsorzebrafishembryoswerefixedinasolutionof2%glutaraldehydewithorwithout4%paraformaldehydein0.1Msodiumcacodylatebufferfollowedbypost-fixationwith1%osmiumtetroxideand1%uranylacetate.Afterdehydrationingradedethanol,cellswereembeddedinEmbed812(ElectronMicroscopySciences)and80nmsectionswerecutusingaLeicaEMUC7microtome.Electronmicrographswereacquiredusingatransmissionelectronmicroscope(Hitachi7650TEM).Forphotoreceptorciliumcountingfromhistologicalysis,semi-thinsections(250nm)cutfromsamplesembeddedforTEMysiswerestainedwithtoluidineblue.Photoreceptoroutersegmentswithintactinnersegmentswerecounted(>100).及实验准备工作，如前期实验结果最好附相应等样品为hTERT-RPE1细胞，三组细胞样品（WT细胞，Centlein-KO-1细胞Centlein-KO-2细胞本对电镜技术并不了解，未做过电镜实验。Fig.4a （EarlystepsinprimaryciliumassemblyrequireEHD1/EHD3-dependentciliaryvesicleformation.）Fig.4G-K （Cep164mediatesvesiculardockingtothemothercentrioleduringearlystepsofciliogenesis.）Fig.7 （TheCP110-i ctingproteinsTalpid3andCep290yoverlaanddistinctrolesinciliaassembly.）Fig.5F（CCDC41isrequiredforciliaryvesicledockingtothemothercentriole.）Fig.6(WDR8isacentriolarsliteandcentriole-associatedproteinthatpromotesciliaryvesicledockingduringFig.6(PrimaryciliogenesisrequiresthedistalappendagecomponentCep123.)公