CN112798378B - Immunofluorescence dry tablet recovery agent and application thereof - Google Patents
Immunofluorescence dry tablet recovery agent and application thereof Download PDFInfo
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- CN112798378B CN112798378B CN202110126830.9A CN202110126830A CN112798378B CN 112798378 B CN112798378 B CN 112798378B CN 202110126830 A CN202110126830 A CN 202110126830A CN 112798378 B CN112798378 B CN 112798378B
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- 238000010166 immunofluorescence Methods 0.000 title claims abstract description 43
- 238000011084 recovery Methods 0.000 title claims abstract description 10
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 29
- 239000008055 phosphate buffer solution Substances 0.000 claims abstract description 20
- 239000000243 solution Substances 0.000 claims abstract description 17
- 229920001213 Polysorbate 20 Polymers 0.000 claims abstract description 9
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 claims abstract description 9
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 claims abstract description 9
- 238000005406 washing Methods 0.000 claims abstract description 8
- 238000002791 soaking Methods 0.000 claims abstract description 4
- 238000002474 experimental method Methods 0.000 abstract description 11
- 238000003018 immunoassay Methods 0.000 abstract description 2
- 230000000694 effects Effects 0.000 description 6
- 238000003556 assay Methods 0.000 description 5
- 238000011534 incubation Methods 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000000465 moulding Methods 0.000 description 1
- 239000003973 paint Substances 0.000 description 1
- 238000013102 re-test Methods 0.000 description 1
- 230000003716 rejuvenation Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/34—Purifying; Cleaning
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
- G01N33/533—Production of labelled immunochemicals with fluorescent label
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Abstract
The invention discloses an immunofluorescence dry tablet recovering agent and application thereof, relating to the fields of immunoassay and biotechnology application, and comprising the following components in parts by weight: 4-6 parts of 5% SDS,4.5-5.5 parts of 0.5% Tween-20, 995-1005 parts of PBS solution, use of the immunofluorescence dry plate recovering agent comprising the steps of: a1: washing the tissue slices or cell samples of the dry slices with PBS solution for 5min each time for 1-2 times; a2: dropwise adding or soaking the immunofluorescence dry-sheet restoration agent on the cleaned tissue slices or cell samples, incubating for 20min by using a shaking table, wherein the temperature is room temperature, and the rotating speed of the shaking table is 60-80 rpm; a3: and (3) washing the incubated tissue slices or cell samples with PBS (phosphate buffer solution) for 5min each time, and finishing the recovery of the immunofluorescence dry slices. The invention can recover the immunofluorescence dry film, can increase the fault tolerance rate of the immunofluorescence experiment, saves the number of samples and saves the experiment time.
Description
Technical Field
The invention relates to the fields of immunoassay and biotechnology application, in particular to an immunofluorescence dry tablet restoring agent and application thereof.
Background
Immunofluorescence is often disposable, i.e., observed by fluorescence microscopy or laser confocal immediately after an experiment. However, in the course of immunofluorescence assay, if there is an operational error, for example, tissue slices or cell sample dry slices are caused by long-term incubation without adding a solution, the background of immunofluorescence is increased, and the final assay result is affected. The prior art can only compensate for errors by re-testing with new tissue sections or cell samples, and often there is no excess sample for making mistakes if the test sample is precious, for example, a tissue sample with infrequent disease or a test sample that requires lengthy molding. For immunofluorescence assay conditions, when studies are needed to cause poor specificity of dry tablets or antibodies, a part of an assay sample is often needed to find suitable assay conditions. This results in an increase in the amount of sample required, prolonging the time of the experiment.
Disclosure of Invention
The invention provides an immunofluorescence dry film restoration agent and application thereof, aiming at the problems in the prior art, the immunofluorescence dry film restoration agent can restore an immunofluorescence dry film, can increase the fault tolerance rate of an immunofluorescence experiment, saves the number of samples and saves the experiment time.
The invention is realized by the following technical scheme: an immunofluorescence dry tablet recovery agent comprises the following components in parts by weight: 4-6 parts of 5-SDS, 4.5-5.5 parts of 0.5% Tween-20, 995-1005 parts of PBS solution.
Further, the paint comprises the following components in parts by weight: 4.59 parts of 5% SDS,5 parts of 0.5% Tween-20, 1000 parts of PBS solution.
The application of the immunofluorescence dry tablet recovering agent comprises the following steps:
a1: washing the tissue slices or cell samples of the dry slices with PBS solution for 5min for 1-2 times;
a2: dropwise adding or soaking the immunofluorescence dry-sheet restoration agent on the cleaned tissue slices or cell samples, incubating for 20min by using a shaking table, wherein the temperature is room temperature, and the rotating speed of the shaking table is 60-80 rpm;
a3: and (3) washing the incubated tissue slices or cell samples with a PBS solution for 5min each time for 3 times to finish the recovery of the immunofluorescence dry slices.
The above includes at least the following advantages:
1. the immunofluorescence dry tablet recovery agent and the application thereof can recover a sealed dry tablet, a primary antibody incubated dry tablet and a secondary antibody incubated dry tablet, and have obvious effects;
2. the immunofluorescence dry film restoration agent and the application thereof can restore the immunofluorescence dry film, can increase the fault tolerance rate of an immunofluorescence experiment, save the number of samples and save the experiment time; meanwhile, experimental conditions are required to be explored for some samples, so that the application of the restoring agent can meet the requirements of dry sample pieces; in addition, some antibodies are less specific and enhanced antibody specificity may also be obtained by the use of the rejuvenating agents of the invention.
Drawings
The accompanying drawings, which are included to provide a further understanding of the embodiments of the invention and are incorporated in and constitute a part of this application, illustrate embodiment(s) of the invention and together with the description serve to explain the principles of the invention. In the drawings:
FIG. 1 is a graph showing the effect of using an immunofluorescence dry plate restoration agent on a dry plate after sample blocking;
FIG. 2 is a graph showing the effect of using an immunofluorescence dry plate restoration agent on a dry plate after a sample primary antibody incubation;
FIG. 3 is a graph showing the effect of using the immunofluorescence dry plate recovering agent on a dry plate after incubation of a sample secondary antibody.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail below with reference to examples and accompanying drawings, and the exemplary embodiments and descriptions thereof are only used for explaining the present invention and are not meant to limit the present invention.
Examples
In one embodiment, the immunofluorescence dry tablet restoration agent comprises the following components in parts by weight: 4-6 parts of 5% SDS,4.5-5.5 parts of 0.5% Tween-20, 995-1005 parts of PBS solution.
In one embodiment, the immunofluorescence dry tablet restoration agent comprises the following components in parts by weight: 4.59 parts of 5-percent SDS,5 parts of 0.5-percent Tween-20, 1000 parts of PBS solution.
In one embodiment, the immunofluorescence dry tablet restoration agent comprises the following components in parts by weight: 4 parts of 5-percent SDS,4.5 parts of 0.5% Tween-20, 995 parts of PBS solution.
In one embodiment, the immunofluorescence dry tablet restoration agent comprises the following components in parts by weight: 6 parts of 5% SDS,5.5 parts of 0.5% Tween-20, 1005 parts of PBS solution.
In one example, 5% SDS,0.5% Tween-20, PBS solution was mixed well to obtain the immunofluorescence dry plate recovering agent.
In one embodiment, the application of the immunofluorescence dry-film restoration agent comprises the following steps:
a1: washing the tissue slices or cell samples of the dry slices with PBS solution for 5min each time for 1-2 times;
a2: dropwise adding or soaking the immunofluorescence dry-sheet restoration agent on the cleaned tissue slices or cell samples, incubating for 20min by using a shaking table, wherein the temperature is room temperature, and the rotating speed of the shaking table is 60-80 rpm;
a3: and (3) washing the incubated tissue slices or cell samples with PBS (phosphate buffer solution) for 5min each time, and finishing the recovery of the immunofluorescence dry slices.
Examples of the experiments
Immunofluorescence dried film recovery effect experiment:
dividing the blocked sample, the primary antibody incubated sample and the secondary antibody incubated sample into three groups, observing the three groups by using a microscope at 200x and 400x respectively, wherein the characteristics of the dry sheet process of the blocked sample, the primary antibody incubated sample and the secondary antibody incubated sample can be seen in figure 1 (a), figure 2 (a) and figure 3 (a), and then treating the three groups by using the immunofluorescence dry sheet restoration agent disclosed by the invention, wherein the treatment method is as shown in an example, and the treatment process is observed by using a microscope at 200x and 400x respectively, and the characteristics of the recovery process of the treated blocked sample, the primary antibody incubated sample and the secondary antibody incubated sample can be seen in figure 1 (b), figure 2 (b) and figure 3 (b).
Experiments show that the immunofluorescence dry tablet restoration agent and the application thereof can restore the closed dry tablet, the primary antibody incubated dry tablet and the secondary antibody incubated dry tablet, and the effect is obvious.
The above-mentioned embodiments are intended to illustrate the objects, technical solutions and advantages of the present invention in further detail, and it should be understood that the above-mentioned embodiments are merely exemplary embodiments of the present invention, and are not intended to limit the scope of the present invention, and any modifications, equivalent substitutions, improvements and the like made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (1)
1. The application of the immunofluorescence dry tablet recovering agent is characterized by comprising the following steps:
the immunofluorescence dry tablet restoration agent comprises the following components in parts by weight: 4-6 parts of 5% SDS,4.5-5.5 parts of 0.5% Tween-20, 995-1005 parts of PBS solution;
a1: washing the tissue slices or cell samples of the dry slices with PBS solution for 5min each time for 1-2 times;
a2: dripping or soaking the immunofluorescence dry-sheet restoration agent on the cleaned tissue slices or cell samples, and incubating for 20min by using a shaking table at room temperature and 60-80 rpm;
a3: and (3) washing the incubated tissue slices or cell samples with PBS (phosphate buffer solution) for 5min each time, and finishing the recovery of the immunofluorescence dry slices.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN202110126830.9A CN112798378B (en) | 2021-01-29 | 2021-01-29 | Immunofluorescence dry tablet recovery agent and application thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110126830.9A CN112798378B (en) | 2021-01-29 | 2021-01-29 | Immunofluorescence dry tablet recovery agent and application thereof |
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| CN112798378A CN112798378A (en) | 2021-05-14 |
| CN112798378B true CN112798378B (en) | 2023-03-24 |
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