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Rapid and economic immunofluorescence method based on cell growing on glass slides

A technology of immunofluorescence and immunofluorescence staining, which is applied in the direction of instruments, measuring devices, scientific instruments, etc., can solve the problems of tissue organs and cell antigen localization detection and visual judgment, etc., and achieve better fixed penetration effect, convenient operation and easy The effect of mastering

Active Publication Date: 2019-02-22
北京科跃中楷生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The method has its own advantages when used in diagnosis and detection, but it cannot be used for antigen localization detection and intuitive judgment of infected tissues, organs and cells

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] An immunofluorescence method based on cell slides, comprising preparation of cell slides and immunofluorescent staining, specifically:

[0042] 1. Preparation of cell slides: PGCs cells were purified and washed, placed in 4% PFA-PBS solution containing 0.01% cysteine ​​on amino-modified glass slides, fixed at room temperature, and then the PFA-PBS solution was sucked off , Cells are fixed while the cell climbing sheet is completed, wherein the modification method of the amino-modified glass climbing sheet is: the glass climbing sheet is placed in a piranha solution containing 0.25% oxalic acid and 0.12% D-cellobiose for processing , the concentrated H in the piranha solution 2 SO 4 and H 2 o 2 The volume ratio of L-cysteine ​​and D-cysteine ​​in cysteine ​​is 1:2.0; the ratio of L-cysteine ​​and D-cysteine ​​is 100:2.8;

[0043] 2. Immunofluorescence staining:

[0044] 1) Blocking: cover the slide with the face down on the blocking solution, and block with 2% antib...

Embodiment 2

[0048] An immunofluorescence method based on cell slides, comprising preparation of cell slides and immunofluorescent staining, specifically:

[0049] 1. Preparation of cell slides: PGCs cells were purified and washed, placed in 4% PFA-PBS solution containing 0.01% cysteine ​​on amino-modified glass slides, fixed at room temperature, and then the PFA-PBS solution was sucked off , complete the cell climbing while the cells are fixed, wherein the modification method of the amino-modified glass climbing piece is: place the glass climbing piece in the piranha solution containing 0.28% oxalic acid and 0.135% D-cellobiose , the concentrated H in the piranha solution 2 SO 4 and H 2 o 2 The volume ratio of cysteine ​​is 1:2.2; the ratio of L-cysteine ​​and D-cysteine ​​in cysteine ​​is 100:3.5;

[0050] 2. Immunofluorescence staining:

[0051] 1) Blocking: cover the slide with the face down on the blocking solution, and block with 2% antibody blocking solution at room temperature...

Embodiment 3

[0055] An immunofluorescence method based on cell slides, comprising preparation of cell slides and immunofluorescent staining, specifically:

[0056] 1. Preparation of cell slides: PGCs cells were purified and washed, placed in 4% PFA-PBS solution containing 0.01% cysteine ​​on amino-modified glass slides, fixed at room temperature, and then the PFA-PBS solution was sucked off , Cells are fixed while the cell climbing sheet is completed, wherein the modification method of the amino-modified glass climbing sheet is: the glass climbing sheet is placed in a piranha solution containing 0.30% oxalic acid and 0.15% D-cellobiose for processing , the concentrated H in the piranha solution 2 SO 4 and H 2 o 2 The volume ratio of L-cysteine ​​and D-cysteine ​​in cysteine ​​is 1:2.5; the ratio of L-cysteine ​​and D-cysteine ​​is 100:4.2;

[0057] 2. Immunofluorescence staining:

[0058] 1) Blocking: cover the slide with the face down on the blocking solution, and block with 2% antib...

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Abstract

The invention provides a rapid and economic immunofluorescence method based on cell growing on glass slides and belongs to the technical field of molecular cell biology. The method comprises preparation of cell growing on glass slides and immunofluorescent staining, wherein the step of preparation of cell growing on glass slides comprises the following steps: purifying PGCs (Primordial Germ Cells), washing, immobilizing the cells on amino modified glass slide to obtain the cell growing on glass slides; and the step of immunofluorescent staining comprises the following steps: placing the glassslide downward on confining liquid for confining; dropwise adding an antibody, incubating, and washing; dropwise adding an anti-fluorescence fading agent on the antibody-labeled slide, absorbing excessive anti-fluorescence fading agent around the slide, and observing fluorescence under fluorescence microscope and shooting. The immunofluorescence method disclosed by the invention is simple and convenient to operate and easy to master and is capable of accurately positioning the antigen position and relative display quantity, and the preparation method of the used cell growing slide has the advantages of being simple in material, cost-saving, excellent in slide effect, uniform in distribution of cells after growing on the slide, convenient to operate, time-saving and the like.

Description

technical field [0001] The invention belongs to the technical field of cell and molecular biology, and in particular relates to an immunofluorescence method based on cell slides. Background technique [0002] Duck Virus Hepatitis (DVH) is an acute and highly fatal infectious disease of ducklings caused by duck hepatitis virus. Opistonus, hepatomegaly and a large number of hemorrhagic spots can be seen in autopsy, which is one of the main diseases that endanger the duck industry. The disease can be preliminarily diagnosed based on epidemiology, typical symptoms and lesions, that is, the onset and death of ducklings, acute onset, rapid death, concentrated death time, and liver swelling with obvious bleeding points or spots. Laboratory tests are still needed to confirm the diagnosis. Currently, laboratory tests mainly include virus isolation and preliminary identification, neutralization test, agar immunodiffusion test, indirect hemagglutination test, dot immunogold filtration...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/533G01N33/569
CPCG01N33/533G01N33/56983
Inventor 曾小敏
Owner 北京科跃中楷生物技术有限公司
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