隨著單細(xì)胞組學(xué)研究的飛速發(fā)展，組織提核技術(shù)逐漸成為單細(xì)胞研究中的必要手段。單核測(cè)序相較單細(xì)胞測(cè)序有著諸多優(yōu)勢(shì)：簡(jiǎn)單快捷的實(shí)驗(yàn)步驟，更少細(xì)胞解離引起的類型偏好，解除細(xì)胞大小對(duì)微流控捕獲效率的偏好，減少解離時(shí)應(yīng)激基因誘導(dǎo)的轉(zhuǎn)錄假象，使細(xì)胞直徑過(guò)大、細(xì)胞形狀不規(guī)則、凍存組織做單細(xì)胞研究成為可能。目前神經(jīng)細(xì)胞[1-3]、心臟[4]、腎臟[5-6]、肝臟[7-8]、脂肪[9-10]等的單細(xì)胞研究均可選用細(xì)胞核進(jìn)行實(shí)驗(yàn)。
然而提核技術(shù)也存在一定問(wèn)題，比如提核后的細(xì)胞核狀態(tài)不好、細(xì)胞核的得率較低、細(xì)胞核易粘連、碎片和雜質(zhì)過(guò)多且難以去除等。
為提高科研服務(wù)水平，優(yōu)化提核技術(shù)，助力單細(xì)胞研究，博奧晶典研究人員經(jīng)過(guò)不斷鉆研，攻克技術(shù)難關(guān)，成功研發(fā)出【動(dòng)物組織細(xì)胞核分離試劑盒】。目前此款試劑盒已成功測(cè)試新鮮組織、凍存組織的多種組織類型，包括小鼠腦、人腦、小鼠心臟、小鼠腎臟、小鼠肝臟、人腦、人卵巢、人肺、豬心臟、肝臟、脂肪等等，數(shù)據(jù)結(jié)果表現(xiàn)優(yōu)秀。
圖1 博奧晶典自研組織細(xì)胞核分離試劑盒
檢驗(yàn)原理
該試劑盒通過(guò)利用表面活性劑破壞組織細(xì)胞的膜和蛋白質(zhì)結(jié)構(gòu)，釋放細(xì)胞核，使之處于游離狀態(tài)，然后通過(guò)密度梯度離心純化細(xì)胞核。得到的純凈細(xì)胞核可適用于單細(xì)胞核轉(zhuǎn)錄組、單細(xì)胞 ATAC-seq 和單細(xì)胞多組學(xué)等實(shí)驗(yàn)。
主要組成成分
表1：博奧晶典自研組織細(xì)胞核分離試劑盒主要組成成分表
部分測(cè)試數(shù)據(jù)展示
圖2 人卵巢 OCT 包埋組織（左上）、豬心臟（右上）、人腦（左下）、小鼠腎臟（右下）提核結(jié)果
圖3 小鼠腦提核數(shù)據(jù)展示（細(xì)胞活率0.12%、結(jié)團(tuán)率5.86%）
圖4 小鼠腦提核樣本的3’單細(xì)胞轉(zhuǎn)錄組小測(cè)結(jié)果（細(xì)胞數(shù)7746，基因中位數(shù)1582）
經(jīng)過(guò)以上數(shù)據(jù)展示，可以發(fā)現(xiàn)博奧晶典自主研發(fā)的核分離試劑盒在各類新鮮組織、凍存組織提核的實(shí)驗(yàn)中表現(xiàn)很好。尤其是從小鼠腦單細(xì)胞核轉(zhuǎn)錄組的實(shí)際小測(cè)數(shù)據(jù)和分析結(jié)果來(lái)看，在測(cè)序飽和度不高的前提下，捕獲細(xì)胞數(shù)、基因中位數(shù)、Fraction Reads in Cell 等重要指標(biāo)非常良好。此外細(xì)胞分群效果良好，星形膠質(zhì)細(xì)胞、少突膠質(zhì)細(xì)胞、小神經(jīng)膠質(zhì)細(xì)胞、興奮性神經(jīng)元、抑制性神經(jīng)元等關(guān)鍵細(xì)胞類型均得到了很好的鑒定，各細(xì)胞類型的數(shù)量和比例符合理論值。
經(jīng)過(guò)上述介紹，相信您對(duì)這款試劑盒已經(jīng)有了一些了解，博奧晶典始終以更高標(biāo)準(zhǔn)的服務(wù)、追求卓越的態(tài)度提供科研服務(wù)，希望博奧晶典自研【動(dòng)物組織細(xì)胞核分離試劑盒】可以助您更好的開(kāi)展單細(xì)胞組學(xué)實(shí)驗(yàn)，為您提供更加準(zhǔn)確的單細(xì)胞組學(xué)方案。
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