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

A detection device and detection method technology, applied in the direction of measuring devices, biochemical cleaning devices, biochemical equipment and methods, etc., can solve the problems of strong subjectivity in manual film reading, heterogeneous reporter genes, and changes in cell structure, etc., to achieve optical The effect of easy control of slice thickness and position, high energy utilization rate, and good imaging signal-to-noise ratio

Active Publication Date: 2019-01-01
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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  • A label-free cell detection device and method based on light sheet illumination
  • A label-free cell detection device and method based on light sheet illumination
  • A label-free cell detection device and method based on light sheet illumination

Examples

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Effect test

Embodiment 1

[0052] A label-free cell detection device and method based on light sheet illumination are used to realize uniformly illuminated light sheets of 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 excitation of light sheets of different thicknesses for samples of different sizes and sizes, and specifically controls the thickness of the light sheets , Limit the excitation area to avoid interference from scattered light from other samples. By controlling the expansion ratio of the beam and the focal length of the cylindrical lens, a light sheet with the required thickness is produced. In this example, light sheets with different thicknesses are realized by converting cylindrical lenses with different focal lengths. The different thicknesses are visualized by rhodamine solution, and the thickness of the l...

Embodiment 2

[0059] In order to verify the sensitivity and accuracy of the present invention for the identification of micron-level particle sizes, polystyrene standard microspheres are 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 scatter diagrams are obtained, and the experimental results and Mie simulation results are compared and analyzed by FFT. The microscopic objective lens selected 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 are both in this range . In this example, the thickness of the light sheet is adjusted to about 13 μm.

[0060] Specific steps:

[0061] (1) Take an appropriate amount of polystyrene microsphere stock so...

Embodiment 3

[0070] A label-free cell detection device and method based on light sheet illumination are used to identify and automatically classify human fibroblast senescent cells and normal cells. In this example, the thickness of the adjusting light sheet is about 50 μm. The cell morphology image and its two-dimensional light scattering pattern were collected on the human fibroblast senescent cells and normal cells that had undergone aging induced by hydrogen peroxide. 55 experimental results were randomly collected for each type. The 110 light scattering images were extracted with feature parameters, and the support vector machine (SVM) algorithm was input as the feature parameters to automatically classify the two types of cells.

[0071] Implementation:

[0072] (1) Prepare the human fibroblast senescent cells and normal cells treated with hydrogen peroxide, configure the cell suspension with PBS buffer, introduce the sample microchamber, control the stage and imaging unit device, and ob...

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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 sub-micron particle size recognition and can be used to classify senescent cells and normal cells, and cell senescence involves a variety of senescent diseases such as malignant tumors And cardiovascular disease. Background technique [0002] Clinically, the detection and classification of cells mainly rely on flow cytometry. The traditional flow cytometer uses a complex optical path system to shape the laser beam into an elliptical beam close to the cell volume to limit the excitation area, reduce interference, and strictly control the sheath flow system to keep the laser beam orthogonal to the sample flow to achieve a single 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 usually need to u...

Claims

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Application Information

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Patent Type & Authority Patents(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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