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Label-free microfluidic cell instrument and method based on light sheet illumination and sheath flow technology

A label-free, microfluidic technology, applied in the direction of instruments, scientific instruments, measuring devices, etc., can solve the problems of being easily affected by incident light and reflected light on the chamber wall, etc., to reduce the possibility of cross-contamination of samples, Overcoming the effect of high cost and simple production process

Active Publication Date: 2018-08-24
SHANDONG UNIV
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  • Description
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Problems solved by technology

Microfluidic light scattering cytometer combines light scattering technology with microfluidic technology, which can realize label-free, fast and non-destructive analysis of cells, but the light scattering detection signal is easily affected by incident light and reflected light from the chamber wall. Especially for weakly scattering samples, it is a very important issue to effectively improve the signal-to-noise ratio of light scattering detection

Method used

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  • Label-free microfluidic cell instrument and method based on light sheet illumination and sheath flow technology
  • Label-free microfluidic cell instrument and method based on light sheet illumination and sheath flow technology
  • Label-free microfluidic cell instrument and method based on light sheet illumination and sheath flow technology

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Embodiment 1

[0063] A label-free microfluidic cytometer based on light sheet illumination and sheath flow technology, through the coupling of light sheet and sheath flow, it realizes single-form arrangement and excitation of fast-flowing cells. By controlling the parameters of the sheath flow generator, different sheath flow effects can be generated to focus the sample flow hydrodynamically, and the thickness of the sample flow varies with different settings of the parameters. In the experiment, the corresponding parameters can be selected according to the size of the sample to be tested, and the thickness of the light sheet suitable for the effect of the sheath flow can be provided to ensure that the particles or cells to be tested in the sample flow can be uniformly and effectively excited, while suppressing the light scattering imaging process interference from background noise. The thickness of the light sheet can be flexibly controlled by using beam expanders with different beam expan...

Embodiment 2

[0090] The invention can realize particle size identification at the submicron resolution level. The simulation database is constructed according to the Mie theory, and the similarity matching based on the Euclidean distance is performed on the collected particle light scattering images to determine the size and refractive index information of the particles. Different from the traditional one-dimensional light scattering method based on relative intensity measurement, the method based on image recognition in the present invention can reduce the influence of incident light and other factors on the stability of detection results during the detection process, and does not require the standard ball calibration process, which simplifies Operational procedure for particle size identification. In this example, a polystyrene microsphere solution with an average diameter of 3.87 μm and a standard deviation of 0.25 μm was selected as the test solution to test and analyze the resolution ...

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Abstract

The invention discloses a label-free microfluidic cell instrument and method based on a light sheet illumination and sheath flow technology, and the method is as follows: a laser beam shaping module generates a laser light sheet to be incident into a sheath flow generation control module, the sheath flow generation control module forms a sheath flow effect by a water dynamic focusing effect, the incident laser light sheet from the laser beam shaping module is coupled with a focused sample stream under the sheath flow effect to excite to-be-tested particles or cells fast flowing in the sample stream to undergo light scattering, and a two-dimensional light scattering image of the to-be-tested particles or cells is captured by an imaging acquisition module and sent to an image processing system for data analysis and result output. A sheath flow machine can be fasted formed by a novel sheath flow technology without complicated micromachining operation; by the coupling of the light sheet illumination technology and the sheath flow technology, the light scattering detection signal to noise ratio can be improved, and label-free fast imaging of the flowing particles or cells can be realized. High-precision particle size recognition in a flow state can be realized by a particle size identification method based on Euclidean distance for matching degree query.

Description

technical field [0001] The invention relates to the fields of biomedical instruments, cell detection technology, microfluidic technology, light sheet technology, etc., and specifically relates to a label-free microfluidic cytometer and method based on light sheet illumination and sheath flow technology. Background technique [0002] Cell recognition and classification are extremely important for clinicopathological analysis and diagnosis. Currently, flow cytometry is one of the main tools for cell detection and analysis. Flow cytometry usually uses sheath flow technology to queue up high-speed cells, and then uses light scattering technology and fluorescent labeling technology to achieve multi-parameter qualitative or quantitative detection and analysis of single cells. Flow cytometry has the advantages of fast detection speed and high specificity by integrating multi-wavelength lasers, multi-color fluorescent labels and multi-channel detection. Its shortcomings are mainly...

Claims

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Application Information

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IPC IPC(8): G01N15/14
CPCG01N15/1434G01N2015/144
Inventor 苏绚涛林梅爱
Owner SHANDONG UNIV
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