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An integrated rapid detection system for real-time fluorescent quantitative analysis of ultra-high-throughput single-cell nucleic acid molecules

A real-time fluorescence quantitative and single-cell nucleic acid technology, which is applied in the measurement/testing of microorganisms, biochemical instruments, laboratory equipment, etc., can solve the problem of low throughput of single-cell analysis, loss of information on cell heterogeneity, and diverse single-cell functions In order to improve detection throughput, realize fast drawing, and reduce calculation

Active Publication Date: 2021-08-24
SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Therefore, the current single-cell analysis throughput is low, and it needs to be combined with a nucleic acid fluorescence real-time quantitative analysis instrument
However, the existing nucleic acid real-time quantitative analysis instruments are based on the analysis of nucleic acid molecules in population cells, which loses the information of cell heterogeneity and the key information of single cell functional diversity.

Method used

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  • An integrated rapid detection system for real-time fluorescent quantitative analysis of ultra-high-throughput single-cell nucleic acid molecules
  • An integrated rapid detection system for real-time fluorescent quantitative analysis of ultra-high-throughput single-cell nucleic acid molecules
  • An integrated rapid detection system for real-time fluorescent quantitative analysis of ultra-high-throughput single-cell nucleic acid molecules

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Embodiment 1

[0079] On the basis of the above examples, refer to Figure 5 and 6 , in this embodiment, the temperature-controlled thermal cycle device 2 includes a mount 20, a temperature-controlled base 21 slidably arranged on the mount 20, and a load for placing the microfluidic chip 4 on the temperature-controlled base 21. stage 22, a heating assembly 23 arranged between the stage 22 and the temperature-controlled base 21, a heat dissipation assembly 25 arranged on the mounting base 20, and a first driving mechanism for driving the temperature-controlled base 21 to slide on the mounting base 20 26. The microfluidic chip 4 is placed on the stage 22 and covered and sealed by a transparent cover.

[0080] Among them, more preferably, the temperature control base 21 is provided with a heat spreader 24, and the heat spreader 24 has a plurality of heat conduction fins to facilitate rapid heat dissipation. A copper plate 27 is arranged on the heat conductor 24, the heating device is a Pelti...

Embodiment 2

[0084] On the basis of the above examples, refer to Figure 1-4 , in this embodiment, the automatic sample loading device 1 includes a sample loading base 10 fixedly connected to the mounting base 20 and an X-axis driving mechanism 11, a Y-axis driving mechanism 12, and a Z-axis driving mechanism 13 arranged on the sample loading base 10. 1. The sample loading mechanical arm 14; the X-axis driving mechanism 11, the Y-axis driving mechanism 12, and the Z-axis driving mechanism 13 are used to realize the movement of the sample loading mechanical arm 14 along the X-axis, Y-axis and Z-axis directions. The sample loading robot 14 is provided with a sample loading needle 140 , which can absorb samples and reagents and add them into the microfluidic chip 4 .

[0085] Further, the X-axis drive mechanism 11 includes an X-axis slide rail 110 arranged on the sample loading base 10, an X-axis driving pulley 111 and an X-axis driven pulley 112, and an X-axis driving pulley 111 and an X-axi...

Embodiment 3

[0090] On the basis of the above examples, refer to Figure 7-10 , in this embodiment, the fluorescence imaging system 3 includes a wide-spectrum light source 30, a switchable fluorescence spectroscopic device 31, and an imaging detector 32. The wide-spectrum light source is more than 30 LED light sources, and the switchable fluorescent spectroscopic device 31 includes a plurality of switchable optical paths The fluorescence spectroscopic module 310;

[0091] The excitation light emitted by the wide-spectrum light source 30 reaches the sample after being reflected by the fluorescence spectroscopic module 310 , and the fluorescence generated by the sample is transmitted through the fluorescence spectroscopic module 310 and enters the imaging detector 32 to realize fluorescence imaging. Of course, the wide-spectrum light source 30 also includes a light-splitting path for transmitting multiple LED light sources to the fluorescent light-splitting module 310 .

[0092] As a prefer...

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Abstract

The invention discloses an integrated rapid detection system for real-time fluorescent quantitative analysis of ultra-high-throughput single-cell nucleic acid molecules, including: a microfluidic chip, an automatic sample adding device, a temperature-controlled thermal cycle device, a fluorescent imaging system, and a data storage and analysis system ; The automatic sample loading device has degrees of freedom in the X-axis, Y-axis and Z-axis directions, and is used to automatically add samples and reagents into the microfluidic chip; Perform analysis, identify positive samples, and draw real-time fluorescence quantitative analysis curves for positive samples. The integrated rapid detection system for real-time fluorescent quantitative analysis of ultra-high-throughput single-cell nucleic acid molecules of the present invention integrates microfluidic chips, automatic sample adding devices, temperature-controlled thermal cycle devices, fluorescent imaging systems, and data storage and analysis systems, which can realize sample The automatic detection processing can capture nucleic acid amplification and real-time fluorescence quantitative curve analysis of single cells on the order of hundreds of thousands or millions.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to an integrated rapid detection system for real-time fluorescent quantitative analysis of single-cell nucleic acid molecules with ultra-high throughput. Background technique [0002] The growth, development, differentiation, aging and pathological changes of the body are all related to the differential expression of genes. The occurrence, development and metastasis of tumors are also related to the mutation and differential expression of genes. Cells in the center of tumor tissue, surrounding cells, cells in metastases, etc. also have different functional characteristics due to differences in genome and transcriptional expression profiles, affecting and Determine the outcome of tumor treatment, etc. [0003] Traditional gene expression research methods usually measure the expression of a gene at the mRNA level. The expression of mRNA level is usually realized by real-time ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M1/38C12M1/34C12M1/00C12Q1/6851
CPCB01L7/5255C12Q1/6851C12Q2531/113C12Q2563/107C12Q2565/629
Inventor 周连群李金泽李龙辉张威张芷齐李敏唐玉国姚佳李传宇郭振
Owner SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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