A fast and economical immunofluorescence method based on cell 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 inability to detect and intuitively judge the antigen localization of tissues, organs and cells, and achieve the saving of related costs and better fixed penetration effect , the effect of simple materials

Active Publication Date: 2022-02-11
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 L-cysteine ​​and D-cysteine ​​in cysteine ​​is 1:2.2; the ratio of L-cysteine ​​and D-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 ...

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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PUM

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Abstract

The invention provides a fast and economical immunofluorescence method based on cell slides, which belongs to the technical field of cell and molecular biology, and includes preparation of cell slides and immunofluorescence staining, wherein the steps of preparing cell slides are: purifying and washing PGCs cells Place the slides on amino-modified glass slides and fix them to obtain cell slides; the steps of immunofluorescence staining are as follows: cover the slides face down on the blocking solution; add antibodies dropwise, incubate and wash; add antibody-labeled slides dropwise Anti-fluorescent fading agent, absorb excess anti-fluorescent fading agent around the slide, then observe the fluorescence and take pictures under a fluorescent microscope. The immunofluorescence method of the present invention is easy to operate, easy to master, can accurately locate the position of the antigen and display the relative amount, and the preparation method of the cell slides used has the advantages of simple materials, cost saving, good slide effect, uniform distribution of cells after slides, It has the advantages of convenient operation and time saving.

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 Patents(China)
IPC IPC(8): G01N33/533G01N33/569
CPCG01N33/533G01N33/56983
Inventor 柳静曾小敏
Owner 北京科跃中楷生物技术有限公司
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