کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
7557460 | 1491339 | 2016 | 15 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
A multi-layer microchip for high-throughput single-cell gene expression profiling
ترجمه فارسی عنوان
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کلمات کلیدی
PBSRT–qPCRfluorescein amiditeMGBROXTCrFAMNTCPDMSCCDlab-on-a-chip - آزمایشگاه روی تراشه، آز-تراشهEDTA - اتیلن دی آمین تترا استیک اسید Ethylenediaminetetraacetic acid - اتیلینیدامین تتراستیک اسیدstandard deviation - انحراف معیارBio-MEMS - بیوگرافی-MEMSCharge-Coupled Device - دستگاه متصل شارژreverse transcription - رونویسی معکوسTemperature coefficient of resistance - ضریب مقاومت دماLoc - محلPhosphate-buffered saline - محلول نمک فسفات با خاصیت بافریquantitative reverse transcription polymerase chain reaction - واکنش زنجیره ای پلی مراز رونویسی معکوسpolymerase chain reaction - واکنش زنجیره ای پلیمرازPCR - واکنش زنجیرهٔ پلیمرازPolydimethylsiloxane - پلیمتیلسیلوکسانMinor groove binder - گیربکس کوچک
موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
چکیده انگلیسی
Microfluidics or Bio-MEMS technology offers significant advantages for performing high-throughput screens and sensitive assays. The ability to correlate single-cell genetic information with cellular phenotypes is of great importance to biology and medicine because it holds the potential to gain insight into disease pathways that is unavailable from ensemble measurements. Previously, we reported two kinds of prototypes for integrated on-chip gene expression profiling at the single-cell level, and the throughput was designed to be 6. In this work, we present a five-layer microfluidic system for parallelized, rapid, quantitative analysis of RNA templates with low abundance at the single-cell level. The microchip contains two multiplexors and one partitioning valve group, and it leverages a matrix (6Â ÃÂ 8) of quantitative reverse transcription polymerase chain reaction (RT-qPCR) units formed by a set of parallel microchannels concurrently controlled by elastomeric pneumatic valves, thereby enabling parallelized handling and processing of biomolecules in a simplified operation procedure. A comprehensive metallic nanofilm with passivation layer is used to run polymerase chain reaction (PCR) temperature cycles. To demonstrate the utility of the approach, artificial synthesized RNA templates (XenoRNA) and mRNA templates from single cells are employed to perform the 48-readout RT-qPCRs. The PCR products are imaged on a fluorescence microscope using a hydrolysis probe/primer set (TaqMan). Fluorescent intensities of passive reference dye and a fluorescein amidite reporter dye are acquired and measured at the end of PCR cycles.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Analytical Biochemistry - Volume 508, 1 September 2016, Pages 1-8
Journal: Analytical Biochemistry - Volume 508, 1 September 2016, Pages 1-8
نویسندگان
Hao Sun,