樣品制備科大組蛋白質(zhì)科學(xué)研究平臺生物成像中心

電鏡成像申請請發(fā)送 掃描電鏡成像請發(fā)送 jianguozha樣品申請請發(fā)送 樣品：申請書登記接收—審核結(jié)果—通知送樣—用戶送樣—制樣完不超過第二個自然周的機(jī)時。同樣，區(qū)域中心內(nèi)課題組樣品優(yōu)先級大樣品技術(shù)服務(wù)時間承諾：對區(qū)域中心內(nèi)用戶，從實(shí)際接收合格樣品到定，報簽字確認(rèn)。用戶在生物成像中心完成科研工作、獲取研究數(shù)據(jù)并據(jù)此相關(guān)成（、著作等，應(yīng)在成果中對生物成像中心予以致謝（詳細(xì)規(guī)定參見生的信息，儀器介紹和樣品信息等，生物成像中心常規(guī)樣品電鏡成 免疫標(biāo)記電鏡成低溫電鏡成 掃描電鏡成熒光顯微 超分辨熒光顯微鏡圖像處 其課題背CentleinCrispr-cas9CentleinhTERT-RPE1本課題組希望通過透射電鏡技術(shù)，觀察野生型和Centlein敲除細(xì)胞中原進(jìn)一步的與纖關(guān)疾病研究有重要意義。泡）希望達(dá)到的效果如下圖例1中所示RepresentativeelectronmicrographsofRPEcellM-centrioles.Scalebar,200nm.參考方法：ElectronmicroscopyprocessinginRPEcells.Briefly,cellsorzebrafishembryoswerefixedinasolutionof2%glutaraldehydewithorwithout4%paraformaldehydein0.1Msodiumcacodylatebufferfollowedbypost-fixationwith1%osmiumtetroxideand1%uranylacetate.Afterdehydrationingradedethanol,cellswereembeddedinEmbed812(ElectronMicroscopySciences)and80nmsectionswerecutusingaLeicaEMUC7microtome.Electronmicrographswereacquiredusingatransmissionelectronmicroscope(Hitachi7650TEM).Forphotoreceptorciliumcountingfromhistologicalysis,semi-thinsections(250nm)cutfromsamplesembeddedforTEMysiswerestainedwithtoluidineblue.Photoreceptoroutersegmentswithintactinnersegmentswerecounted(>100).及實(shí)驗(yàn)準(zhǔn)備工作，如前期實(shí)驗(yàn)結(jié)果最好附相應(yīng)等樣品為hTERT-RPE1細(xì)胞，三組細(xì)胞樣品（WT細(xì)胞，Centlein-KO-1細(xì)胞Centlein-KO-2細(xì)胞本對電鏡技術(shù)并不了解，未做過電鏡實(shí)驗(yàn)。Fig.4a （EarlystepsinprimaryciliumassemblyrequireEHD1/EHD3-dependentciliaryvesicleformation.）Fig.4G-K （Cep164mediatesvesiculardockingtothemothercentrioleduringearlystepsofciliogenesis.）Fig.7 （TheCP110-i ctingproteinsTalpid3andCep290yoverlaanddistinctrolesinciliaassembly.）Fig.5F（CCDC41isrequiredforciliaryvesicledockingtothemothercentriole.）Fig.6(WDR8isacentriolarsliteandcentriole-associatedproteinthatpromotesciliaryvesicledockingduringFig.6(PrimaryciliogenesisrequiresthedistalappendagecomponentCep123.)公