據(jù)美國(guó)物理學(xué)家組織網(wǎng)近日?qǐng)?bào)道,，最近,，加拿大英屬哥倫比亞大學(xué)與英屬哥倫比亞癌癥研究所、轉(zhuǎn)化與應(yīng)用基因組學(xué)中心合作,，開(kāi)發(fā)出一種硅酮材料的芯片實(shí)驗(yàn)室技術(shù),，能讓每個(gè)細(xì)胞像彈球機(jī)里的球一樣各就各位，然后進(jìn)行基因檢測(cè),。這種“單細(xì)胞基因分析”技術(shù)使基因檢測(cè)更加靈敏迅速,，有助于腫瘤分析和臨床疾病的診斷。本周出版的美國(guó)《國(guó)家科學(xué)院院刊》對(duì)該芯片實(shí)驗(yàn)室進(jìn)行了詳細(xì)介紹,。
這種芯片實(shí)驗(yàn)室大小跟一個(gè)9伏電池相當(dāng),，能同時(shí)分析300個(gè)細(xì)胞,。研究人員設(shè)計(jì)了一種路線，用液體載運(yùn)細(xì)胞通過(guò)顯微管道和一個(gè)個(gè)小閥門,，當(dāng)細(xì)胞挨個(gè)進(jìn)入各自的小空位時(shí),，它們的RNA就會(huì)被提取出來(lái)，經(jīng)過(guò)復(fù)制用于進(jìn)一步分析,。
標(biāo)準(zhǔn)基因檢測(cè)要求使用大量細(xì)胞,，才能得出由上千萬(wàn)個(gè)不同細(xì)胞平均化以后的“綜合圖像”，這會(huì)掩蓋細(xì)胞的真實(shí)屬性和它們之間的相互作用,。“這就好比用混合水果慕絲來(lái)研究草莓和樹(shù)莓為什么不一樣,。”領(lǐng)導(dǎo)該研究的高通量生物中心副教授卡爾·漢森介紹說(shuō)，而單細(xì)胞分析正在成為基因研究中的黃金手段,，因?yàn)榧词故菑耐荒[瘤組織中采集的樣本,，也包含了正常細(xì)胞和多種癌細(xì)胞類型，而單細(xì)胞分析顯出極微小的差異,。
此外,，這種芯片實(shí)驗(yàn)室?guī)缀鯇⑺屑?xì)胞分析過(guò)程整合在了一起，不僅能分離細(xì)胞,，還能用化學(xué)試劑將細(xì)胞混合起來(lái),，通過(guò)檢測(cè)反應(yīng)過(guò)程中的熒光發(fā)射獲得它們的基因編碼。所有這些都能在芯片上完成,，不僅操作簡(jiǎn)單,，而且成本效益高。(生物谷 Bioon.com)
doi:10.1073/pnas.1019446108
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PMID:
High-throughput microfluidic single-cell RT-qPCR
White, Adam K.; VanInsberghe, Michael; Petriv, Oleh I.; Hamidi, Mani; Sikorski, Darek; Marra, Marco A.; Piret, James; Aparicio, Samuel; Hansen, Carl L.
A long-sought milestone in microfluidics research has been the development of integrated technology for scalable analysisof transcription in single cells. Here we present a fully integrated microfluidic device capable of performing high-precisionRT-qPCR measurements of gene expression from hundreds of single cells per run. Our device executes all steps of single-cellprocessing, including cell capture, cell lysis, reverse transcription, and quantitative PCR. In addition to higher throughputand reduced cost, we show that nanoliter volume processing reduced measurement noise, increased sensitivity, and providedsingle nucleotide specificity. We apply this technology to 3,300 single-cell measurements of (i) miRNA expression in K562 cells, (ii) coregulation of a miRNA and one of its target transcripts during differentiation in embryonic stem cells, and (iii) single nucleotide variant detection in primary lobular breast cancer cells. The core functionality established here providesthe foundation from which a variety of on-chip single-cell transcription analyses will be developed.
