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相关概念视频

Real Time RT-PCR02:57

Real Time RT-PCR

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Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
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Simple Bulk Readout of Digital Nucleic Acid Quantification Assays
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基于芯片的数字PCR与大场一次性成像,用于高灵敏度核酸量化.

Jinrong Shen1, Jingxing Fan1, Zhehao Zhao2

  • 1State Key Laboratory of ASIC and System, Fudan University, Shanghai, 200433, China; School of Microelectronics, Fudan University, Shanghai, 200433, China.

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概括

我们开发了一种基于芯片的数字聚合酶连锁反应 (cdPCR) 装置,配备了一种新的大场成像系统. 该系统能够快速,一次性获取图像,改善dPCR分析中的绝对量化.

关键词:
绝对量化的绝对量化.基于芯片的数字PCR.光图像分析 光图像分析高均性的照明系统.肺癌的DNA 肺癌的DNA一次性成像成像技术

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科学领域:

  • 生物技术是生物技术.
  • 分析化学 分析化学
  • 医学诊断 医学诊断 医学诊断

背景情况:

  • 数字聚合酶连锁反应 (dPCR) 为绝对量化提供了高灵敏度.
  • 传统的dPCR成像系统由于图像拼接而面临速度和准确性的限制.

研究的目的:

  • 开发一种基于芯片的增强型dPCR (cdPCR) 装置,具有大场成像系统.
  • 克服传统dPCR成像的局限性,提高绝对检测极限和精度.

主要方法:

  • 设计了一个cdPCR装置,拥有21,384个微反应室.
  • 开发了一种大场光成像系统 (28毫米×20毫米) 用于一次性图像采集.
  • 实现了科勒照明和图像处理模块,以提高图像质量和精度.

主要成果:

  • 在1秒内实现了整个cdPCR芯片的一次性光图像采集.
  • 证明了90%的照明均性,用于高质量的成像.
  • 用肺癌DNA验证了系统,显示测量值和理论值之间的良好相关性 (R2 > 0.9997).

结论:

  • 开发的系统为cdPCR光成像处理提供了一个自动化,快速和准确的平台.
  • 消除图像拼接错误可以提高绝对量化的精度.
  • 该系统推进dPCR分析,以改善诊断应用.