利用微流體PCR進行準確可靠的單細胞基因表達分析
· 達到更準確地檢測單一細胞基因表達譜差異性
· 避免測量樣品中所有細胞的平均值的壞處
· 鑒定以前不能分辨的細胞亞群和分解新的調(diào)控網(wǎng)絡(luò)
在名義上均一的細胞群中，單一細胞在尺寸、蛋白質(zhì)水平和mRNA表達轉(zhuǎn)錄上都存在差異，所以默認你的樣品中每一個細胞都表現(xiàn)得完全一致是一種危險的賭博，測量集中到一起的多個細胞的平均值會掩蓋細胞之間基因表達的顯著差異。在看起來均一的細胞群中辨別細胞間的差異對于促進干細胞研究、理解癌細胞、鑒定免疫反應(yīng)、研究生物治療的有效性、發(fā)現(xiàn)退行性神經(jīng)疾病的機理等方面都有至關(guān)重要的意義。
遺傳學(xué)家和臨床工作者一直在尋找一套完整的實驗流程來檢測分辨單一細胞，根據(jù)它們獨特的基因組和轉(zhuǎn)錄組進行分群，同時將技術(shù)的噪音最小化。
Fluidigm基于微流體技術(shù)開發(fā)了一種全新的單細胞基因表達檢測方法，可以使你快速可靠地分離、處理、對單一細胞的基因組進行分析。我們的儀器、芯片、試劑、軟件、試劑盒組成的一套產(chǎn)品線家族,利用Fluidigm設(shè)計的產(chǎn)品能幫助您只需采用一種技術(shù)即可完成細胞分離提取、逆轉(zhuǎn)錄和預(yù)擴增、到最終監(jiān)測和分析細胞活動等一系列實驗流程。
Fluidigm單細胞流程可以在幾個小時內(nèi)測試數(shù)以百計的單細胞中數(shù)百個基因的表達，這種實驗用傳統(tǒng)的系統(tǒng)通常要用幾天的時間。它提供簡便的流程和靈活的試劑組合。利用自動化的流程、隨時可用的試劑和優(yōu)化的芯片，您可以免除繁重的加樣和樣品混合步驟而達到“加樣走人”的生產(chǎn)率。
Fluidigm的微流體芯片結(jié)構(gòu)可以利用比傳統(tǒng)體系顯著少量的試劑自動完成樣品與試劑、引物探針的混合并建立數(shù)以千計的PCR反應(yīng)體系。每個微流體芯片可以生成相當于24塊標準384-孔板數(shù)據(jù)量。
新的Fluidigm單細胞流程為研究細胞分化、測量單一細胞對特定刺激的反應(yīng)、證實重要疾病生物標志物、驗證RNA干擾沉默基因表達和實行候選藥物篩選等開啟了新的大門。
該流程的技術(shù)平臺為Fluidigm新推出的C1單細胞自動制備系統(tǒng)和已在單細胞基因表達分析中廣泛被采用的BioMark™ HD微流體PCR系統(tǒng)組成。
C1™單細胞自動制備系統(tǒng)
1 小時內(nèi)即可分離制備96個單細胞
BIOMARK™ HD系統(tǒng)
30分鐘內(nèi)可進行9,216個 qPCR反應(yīng)
C1單細胞自動制備系統(tǒng)結(jié)合BioMark™ HD系統(tǒng)持續(xù)有效地優(yōu)化了基因表達分析，支持多達96個單一細胞的96個轉(zhuǎn)錄子的分析。顯著地提高了效率并進一步加速了單細胞基因表達譜的研究。
整個流程可以讓您輕松實現(xiàn)如下步驟：
· 捕獲—組細胞只需一步加樣即能迅速地分離到96個獨立的反應(yīng)倉完成制備
· 確認—質(zhì)控節(jié)點確認捕獲細胞數(shù)量及分辨活細胞和死細胞
· 裂解—快速直接的細胞裂解方法可節(jié)約時間和費用且不需要RNA純化步驟
· 逆轉(zhuǎn)錄和預(yù)擴增—cDNA合成及特異目的片段擴增在一個樣品中進行且不需要試劑混合及樣品轉(zhuǎn)移
· 收獲—所有擴增產(chǎn)物被集中到一起、收集和轉(zhuǎn)移到BioMark™ HD系統(tǒng)進行實時PCR分析
· 轉(zhuǎn)移—在C1系統(tǒng)上預(yù)擴增的產(chǎn)物被稀釋加樣到動態(tài)芯片IFC上
· 定量PCR-將動態(tài)芯片IFC放到BioMark™ HD系統(tǒng)上進行PCR反應(yīng)和數(shù)據(jù)采集等
· 分析—用戶友好界面的軟件包可實時查看擴增曲線、獲取彩色色熱圖及Ct數(shù)據(jù)等
使用該平臺發(fā)表的最新文章
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