Label-free cell detection device and method based on light sheet illumination

A detection device and detection method technology, applied in measurement devices, biochemical cleaning devices, biochemical equipment and methods, etc., can solve the problems of strong subjectivity of manual reading, uneven reporter genes, complex process steps, etc. The effect of easy control of sheet thickness and position, efficient energy utilization, and low cost

Active Publication Date: 2017-03-22
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Flow cytometry measurements can more clearly distinguish senescent cells, but random insertion of reporter genes will cause additional differential expression, resulting in heterogeneous reporter genes and affecting detection results
All in all, the above methods are easy to detect and have high accuracy, but the process steps are complex, time-consuming, and costly. At the same time, manual reading is highly subjective and time-consuming, and the cell labeling and staining processes may also change. Cell structure, affecting experimental results

Method used

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  • Label-free cell detection device and method based on light sheet illumination
  • Label-free cell detection device and method based on light sheet illumination
  • Label-free cell detection device and method based on light sheet illumination

Examples

Experimental program
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Effect test

Embodiment 1

[0053] A label-free cell detection device and method based on light sheet illumination is used to realize uniform illumination light sheets with different thicknesses. In light scattering imaging, in order to reduce the background noise in the imaging process and at the same time ensure that the sample to be tested is uniformly illuminated, the present invention provides different thicknesses of light sheet excitation for samples of different sizes and scales, and controls the thickness of the light sheet in a targeted manner. , to limit the excitation area and avoid interference from scattered light from other samples. A light sheet of required thickness is produced by controlling the beam expansion factor of the beam and the focal length of the cylindrical lens. In this example, light sheets with different thicknesses are realized by converting cylindrical lenses with different focal lengths. The different thicknesses are visualized through a rhodamine solution and the ligh...

Embodiment 2

[0060] In order to verify the sensitivity and accuracy of the present invention for particle size identification at the micron level, polystyrene standard microspheres were used for experimental verification and device calibration. In the present invention, standard microspheres with particle diameters of 3.87 μm and 4.19 μm are selected as imaging samples, and their microscopic images and two-dimensional scattering diagrams are obtained, and the experimental results are compared with the Mie simulation results by FFT analysis. The selected microscopic objective lens in the present invention is a 20 times objective lens with a numerical aperture of 0.4, and its corresponding detectable angle range is 72.5°-107.5°, so in the Mie simulation, the scattering polar angle and azimuth angle values ​​are in this range . In this example, the thickness of the optical sheet is adjusted to be about 13 μm.

[0061] Specific steps:

[0062] (1) Take an appropriate amount of polystyrene mi...

Embodiment 3

[0071] Cell identification and automatic classification of human fibroblast senescent cells and normal cells are performed using a label-free cell detection device and method based on light sheet illumination. In this example, the thickness of the light sheet is adjusted to be about 50 μm. Cell morphology images and two-dimensional light scattering patterns were collected from human fibroblast senescent cells and normal cells induced by hydrogen peroxide aging, and 55 experimental results were randomly collected for each type. The feature parameters of the obtained 110 light scattering images were extracted, and the support vector machine (SVM) algorithm was input as the feature parameters to automatically classify the two types of cells.

[0072] Implementation:

[0073](1) Prepare human fibroblast senescent cells and normal cells treated with hydrogen peroxide, configure cell suspension with PBS buffer, import the sample into the microchamber, control the displacement stage...

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Abstract

The invention discloses a label-free cell detection device and method based on light sheet illumination. The method includes the steps that a light sheet generation unit is used for shaping a laser beam into a uniform illumination light sheet with the micron-order thickness through a cylinder lens; the light sheet excites particles or a cell suspension placed in a sample micro-cavity chamber controlled by a precise displacement table to move; a form microscopic image and a two-dimensional light scattering pattern of a single particle or cell are recorded by a detector under the focusing mode and the defocusing mode through an objective lens; and a result is input into an imaging analysis system for image processing and recognizing and classifying. By means of the light sheet illumination method, a stimulation area can be effectively limited, background interference in light scattering imaging is inhibited, and effective stimulation of the single particle or cell and particle size discrimination of the sub-micron resolution ratio level are realized; and a light sheet illumination stimulation two-dimensional light scattering technology can avoid complex dyeing operation and a fluorescence signal detection process, and label-free detection and classification are carried out on aging cells. The label-free cell detection device and method based on light sheet illumination are high in applicability and capable of being popularized.

Description

technical field [0001] The invention relates to a label-free cell detection device and method based on light sheet illumination, which can realize submicron particle size identification and can be used to classify aging cells and normal cells, and cell aging involves various aging diseases such as malignant tumors and cardiovascular disease, etc. Background technique [0002] The clinical detection and classification of cells mainly relies on flow cytometry. Traditional flow cytometers use complex optical path systems to shape the laser beam into an elliptical beam that is close to the volume of the cell to limit the excitation area, reduce interference, and strictly control the sheath flow system to keep the laser beam orthogonal to the sample flow, so as to achieve a single For the purpose of sample signal detection and acquisition, the control process is complicated and the cost is high. In the detection and classification of biological cells, traditional cytometers usu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/34G01N15/14
CPCG01N15/1434G01N2015/0065G01N2015/1006G01N2015/144G01N2015/149G01N2015/1493
Inventor 苏绚涛林梅爱刘巧谯旭
Owner SHANDONG UNIV
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