CN114181995A - Sample pre-hybridization pretreatment reagent and use method thereof - Google Patents
Sample pre-hybridization pretreatment reagent and use method thereof Download PDFInfo
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- CN114181995A CN114181995A CN202111505159.5A CN202111505159A CN114181995A CN 114181995 A CN114181995 A CN 114181995A CN 202111505159 A CN202111505159 A CN 202111505159A CN 114181995 A CN114181995 A CN 114181995A
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- 238000009396 hybridization Methods 0.000 title claims abstract description 28
- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 25
- 238000000034 method Methods 0.000 title claims abstract description 18
- 239000000243 solution Substances 0.000 claims abstract description 46
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 35
- 102000057297 Pepsin A Human genes 0.000 claims abstract description 26
- 108090000284 Pepsin A Proteins 0.000 claims abstract description 26
- 229940111202 pepsin Drugs 0.000 claims abstract description 26
- 239000008213 purified water Substances 0.000 claims abstract description 23
- 239000003480 eluent Substances 0.000 claims abstract description 20
- 102000004190 Enzymes Human genes 0.000 claims abstract description 18
- 108090000790 Enzymes Proteins 0.000 claims abstract description 18
- 229940088598 enzyme Drugs 0.000 claims abstract description 18
- 230000018044 dehydration Effects 0.000 claims abstract description 16
- 238000006297 dehydration reaction Methods 0.000 claims abstract description 16
- 238000002791 soaking Methods 0.000 claims abstract description 16
- 239000007853 buffer solution Substances 0.000 claims abstract description 14
- 239000007788 liquid Substances 0.000 claims abstract description 14
- 238000001035 drying Methods 0.000 claims abstract description 11
- 239000012224 working solution Substances 0.000 claims abstract description 9
- 230000029087 digestion Effects 0.000 claims abstract description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 27
- 239000011521 glass Substances 0.000 claims description 21
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 18
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 18
- 238000005303 weighing Methods 0.000 claims description 12
- 238000003756 stirring Methods 0.000 claims description 9
- 235000019441 ethanol Nutrition 0.000 claims description 7
- 102000004169 proteins and genes Human genes 0.000 claims description 7
- 108090000623 proteins and genes Proteins 0.000 claims description 7
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 claims description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 6
- 238000010828 elution Methods 0.000 claims description 6
- 230000008014 freezing Effects 0.000 claims description 6
- 238000007710 freezing Methods 0.000 claims description 6
- 230000002496 gastric effect Effects 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 6
- 238000002360 preparation method Methods 0.000 claims description 6
- -1 sodium cyanogen sulfate Chemical compound 0.000 claims description 6
- 239000000872 buffer Substances 0.000 claims description 4
- 239000006059 cover glass Substances 0.000 claims description 3
- 238000007865 diluting Methods 0.000 claims description 3
- 239000003292 glue Substances 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 238000007789 sealing Methods 0.000 claims description 3
- 239000011780 sodium chloride Substances 0.000 claims description 3
- 239000001509 sodium citrate Substances 0.000 claims description 3
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 claims description 3
- 229940038773 trisodium citrate Drugs 0.000 claims description 3
- 239000000523 sample Substances 0.000 abstract description 26
- 238000001514 detection method Methods 0.000 abstract description 4
- 230000000694 effects Effects 0.000 abstract description 4
- 210000001519 tissue Anatomy 0.000 abstract description 4
- 210000004027 cell Anatomy 0.000 abstract description 3
- 210000003855 cell nucleus Anatomy 0.000 abstract description 2
- 230000035699 permeability Effects 0.000 abstract description 2
- 208000005156 Dehydration Diseases 0.000 abstract 2
- 239000008096 xylene Substances 0.000 description 4
- 150000007523 nucleic acids Chemical class 0.000 description 3
- 238000007901 in situ hybridization Methods 0.000 description 2
- 102000039446 nucleic acids Human genes 0.000 description 2
- 108020004707 nucleic acids Proteins 0.000 description 2
- 230000004075 alteration Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 239000002906 medical waste Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6841—In situ hybridisation
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Abstract
The invention discloses a pretreatment reagent before sample hybridization and a use method thereof, wherein pretreatment liquid, enzyme buffer solution, pepsin and eluent are used for sequentially carrying out the procedures of chip baking, dewaxing, dehydration, pretreatment soaking, solution removal, digestion by pepsin working solution, purified water soaking, secondary dehydration and drying on a sample, so that the permeability of sample tissues can be fully improved, the probe and target DNA can be ensured to be fully contacted and hybridized when being hybridized with the probe, after counterstaining by counterstaining liquid, a fluorescent signal is observed under a matched fluorescent microscope, the effect can be that cells are dispersed, cell nucleuses are complete, the fluorescent signal identified by naked eyes can be emitted, and the improvement of the detection effect is facilitated.
Description
Technical Field
The invention relates to the technical field of sample treatment, in particular to a pretreatment reagent before sample hybridization and a use method thereof.
Background
Paraffin-embedded tissue sections (FFPE samples) are commonly used for in situ hybridization detection, which refers to a process of hybridizing specifically labeled nucleic acids of known sequence as probes with nucleic acids in cells or tissue sections, thereby precisely and quantitatively locating specific nucleic acid sequences.
Since the target sequence (DNA) of the paraffin-embedded sample to be detected is located in the nucleus and is encapsulated by proteins, a pretreatment is required before the sample is subjected to in situ hybridization detection.
Disclosure of Invention
The invention aims to provide a pretreatment reagent before sample hybridization and a using method thereof.
In order to achieve the purpose, the invention provides the following technical scheme: a pre-hybridization pretreatment reagent for a sample, characterized in that: the method is characterized in that: the pretreatment reagent consists of pretreatment liquid, enzyme buffer liquid, pepsin and eluent: (1) weighing 81.07-81.1g of sodium cyanogen sulfate, adding purified water into a glass bottle, fixing the volume to 1L, and stirring until the sodium cyanogen sulfate is completely dissolved to obtain a pretreatment solution;
(2) enzyme buffer solution: weighing 5.5-5.6ml of concentrated hydrochloric acid in a glass bottle, adding purified water to dilute to 1000ml, and mixing uniformly to obtain an enzyme buffer solution;
(3) pepsin: weighing 140 and 141mg of gastric protein powder, pouring the gastric protein powder into a freezing tube, and tightly covering the freezing tube with a cover to obtain the pepsin;
(4) eluent: weighing 175-175.3g of sodium chloride and 88-88.2g of trisodium citrate, adding into a proper container, adding purified water to a constant volume of 1L, stirring until the solution is completely dissolved, and adjusting the pH value to 5.3 +/-0.1 by using 1mol/L hydrochloric acid or 1mol/L sodium hydroxide solution to obtain a 20 XSSC solution;
measuring the obtained 20 XSSC solution into 100ml and 3ml of NP-40, adding into a proper container, adding purified water to a constant volume of 1L, stirring uniformly, and adjusting the pH value to 7.0-7.5 by using 11mol/L hydrochloric acid or 1mol/L sodium hydroxide solution to obtain the eluent.
A method for using a pretreatment reagent before sample hybridization is characterized in that: comprises the steps of reagent preparation, sample treatment and elution after hybridization.
The reagent preparation comprises the following steps:
s1: pouring 50mL of pretreatment liquid into a dye vat, placing the dye vat in a water bath kettle at 80 +/-1 ℃, and keeping the temperature of the solution in the dye vat at 80 +/-1 ℃ for later use;
s2: pouring 50mL of enzyme buffer solution into a dye vat, placing the dye vat in a water bath kettle at the temperature of 37 +/-1 ℃, keeping the temperature of the solution in the dye vat at 37 +/-1 ℃ for later use, adding 140mg of pepsin into the enzyme buffer solution before dewaxing a sample, and fully dissolving to obtain a pepsin working solution;
s3: preparing 2 dye vats, adding 50mL of eluent respectively, and placing the dye vats in water bath pots at normal temperature and 73 +/-1 ℃ for later use;
s4: respectively measuring 700mL and 850mL of absolute ethyl alcohol, respectively diluting to 1000mL by using purified water, and sealing and storing at room temperature for later use.
The sample processing comprises the following steps:
s1 baking slices: slightly drying the slices in the air, and baking the slices on a 65-70 baking sheet machine for 3.8-4h for later use;
s2 dewaxing: at room temperature, the glass slide is immersed into xylene or a xylene substitute for dewaxing twice, and each time lasts for 10-15 minutes;
s3 dehydration: after dewaxing is finished, soaking the glass slide into absolute ethyl alcohol for dehydration twice at room temperature for 5 minutes respectively; taking out the slide, and removing residual solution on the slide;
s4 pretreatment: immersing the slide into the pretreatment solution with the temperature of 80 +/-1 ℃ prepared in the first step for treatment for 10 minutes;
s5 removal of solution: taking out the slide, immersing the slide in purified water for 1 minute, and removing residual solution on the slide after taking out;
s6 digestion: placing the glass slide in a pepsin working solution, and digesting for 10-60 minutes at 37 ℃;
s7 soaking: after digestion, the slides were soaked in purified water for 1 minute;
s8 dehydration: after soaking, respectively soaking the glass slides into 70 percent and 85 percent ethanol solutions obtained in the step one and 100 percent ethanol for gradient dehydration for 1 minute respectively;
s9 drying: and (4) taking out the slide after dehydration, and drying at room temperature for later use, thus obtaining the hybridization.
The post-hybridization elution comprises the following steps:
s1: after the end, taking out the slide from the hybridization instrument, and removing the mounting glue by using tweezers;
s2: at room temperature, immersing the glass slide into the eluent which is arranged in the first step and is reserved at the normal temperature for 5 minutes, and removing the cover glass;
s3: taking out the slide, putting the slide into another eluent which is arranged in the water bath kettle in the step one and is prepared for standby and preheated to 73 +/-1 ℃, and rinsing for 2 minutes;
s4: and taking out the slide, removing residual liquid, and airing in a dark place for later use.
The dye vat adopts a Cooprine dye vat.
Compared with the prior art, the invention has the following beneficial effects:
the pretreatment solution, the enzyme buffer solution, the pepsin and the eluent are used for sequentially baking the sample, dewaxing, dehydrating, pretreating and soaking, removing the solution, digesting by using the pepsin working solution, soaking by using purified water, dehydrating again and drying, the permeability of sample tissues can be fully improved, the probe and target DNA can be fully contacted and hybridized when hybridized with the probe, after counterstaining by using the counterstaining solution, a fluorescent signal is observed under a matched fluorescent microscope, the effect can be that cells are dispersed, cell nucleuses are complete, the fluorescent signal identified by naked eyes can be emitted, and the detection effect is favorably improved.
Detailed Description
The technical solutions in the embodiments of the present invention are clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In order to achieve the purpose, the invention provides the following technical scheme: a pre-hybridization pretreatment reagent for a sample, characterized in that: the method is characterized in that: the pretreatment reagent consists of pretreatment liquid, enzyme buffer liquid, pepsin and eluent: (1) weighing 81.07-81.1g of sodium cyanogen sulfate, adding purified water into a glass bottle, fixing the volume to 1L, and stirring until the sodium cyanogen sulfate is completely dissolved to obtain a pretreatment solution;
(2) enzyme buffer solution: weighing 5.5-5.6ml of concentrated hydrochloric acid in a glass bottle, adding purified water to dilute to 1000ml, and mixing uniformly to obtain an enzyme buffer solution;
(3) pepsin: weighing 140 and 141mg of gastric protein powder, pouring the gastric protein powder into a freezing tube, and tightly covering the freezing tube with a cover to obtain the pepsin;
(4) eluent: weighing 175-175.3g of sodium chloride and 88-88.2g of trisodium citrate, adding into a proper container, adding purified water to a constant volume of 1L, stirring until the solution is completely dissolved, and adjusting the pH value to 5.3 +/-0.1 by using 1mol/L hydrochloric acid or 1mol/L sodium hydroxide solution to obtain a 20 XSSC solution;
measuring the obtained 20 XSSC solution into 100ml and 3ml of NP-40, adding into a proper container, adding purified water to a constant volume of 1L, stirring uniformly, and adjusting the pH value to 7.0-7.5 by using 11mol/L hydrochloric acid or 1mol/L sodium hydroxide solution to obtain the eluent.
A method for using a pretreatment reagent before sample hybridization is characterized in that: comprises the steps of reagent preparation, sample treatment and elution after hybridization.
The reagent preparation comprises the following steps:
s1: pouring 50mL of pretreatment liquid into a dye vat, placing the dye vat in a water bath kettle at 80 +/-1 ℃, and keeping the temperature of the solution in the dye vat at 80 +/-1 ℃ for later use;
s2: pouring 50mL of enzyme buffer solution into a dye vat, placing the dye vat in a water bath kettle at the temperature of 37 +/-1 ℃, keeping the temperature of the solution in the dye vat at 37 +/-1 ℃ for later use, adding 140mg of pepsin into the enzyme buffer solution before dewaxing a sample, and fully dissolving to obtain a pepsin working solution;
s3: preparing 2 dye vats, adding 50mL of eluent respectively, and placing the dye vats in water bath pots at normal temperature and 73 +/-1 ℃ for later use;
s4: respectively measuring 700mL and 850mL of absolute ethyl alcohol, respectively diluting to 1000mL by using purified water, and sealing and storing at room temperature for later use.
The sample processing comprises the following steps:
s1 baking slices: slightly drying the slices in the air, and baking the slices on a 65-70 baking sheet machine for 3.8-4h for later use;
s2 dewaxing: at room temperature, the glass slide is immersed into xylene or a xylene substitute for dewaxing twice, and each time lasts for 10-15 minutes;
s3 dehydration: after dewaxing is finished, soaking the glass slide into absolute ethyl alcohol for dehydration twice at room temperature for 5 minutes respectively; taking out the slide, and removing residual solution on the slide;
s4 pretreatment: immersing the slide into the pretreatment solution with the temperature of 80 +/-1 ℃ prepared in the first step for treatment for 10 minutes;
s5 removal of solution: taking out the slide, immersing the slide in purified water for 1 minute, and removing residual solution on the slide after taking out;
s6 digestion: placing the glass slide in a pepsin working solution, and digesting for 10-60 minutes at 37 ℃;
s7 soaking: after digestion, the slides were soaked in purified water for 1 minute;
s8 dehydration: after soaking, respectively soaking the glass slides into 70 percent and 85 percent ethanol solutions obtained in the step one and 100 percent ethanol for gradient dehydration for 1 minute respectively;
s9 drying: and (4) taking out the slide after dehydration, and drying at room temperature for later use, thus obtaining the hybridization.
The post-hybridization elution comprises the following steps:
s1: after the end, taking out the slide from the hybridization instrument, and removing the mounting glue by using tweezers;
s2: at room temperature, immersing the glass slide into the eluent which is arranged in the first step and is reserved at the normal temperature for 5 minutes, and removing the cover glass;
s3: taking out the slide, putting the slide into another eluent which is arranged in the water bath kettle in the step one and is prepared for standby and preheated to 73 +/-1 ℃, and rinsing for 2 minutes;
s4: and taking out the slide, removing residual liquid, and airing in a dark place for later use.
The dye vat adopts a Cooprine dye vat.
The pepsin in the working process is stored at the temperature of 2-8 ℃, and the pepsin working solution consisting of the pepsin needs to be prepared for use.
The pretreatment solution and the eluate should be properly treated as medical waste after one day of use.
The 70% and 85% ethanol solutions were stopped after 20 slides were treated or turbidity and contamination occurred.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (7)
1. A pre-hybridization pretreatment reagent for a sample, comprising: the pretreatment reagent consists of pretreatment liquid, enzyme buffer liquid, pepsin and eluent: (1) weighing 81.07-81.1g of sodium cyanogen sulfate, adding purified water into a glass bottle, fixing the volume to 1L, and stirring until the sodium cyanogen sulfate is completely dissolved to obtain a pretreatment solution;
(2) enzyme buffer solution: weighing 5.5-5.6ml of concentrated hydrochloric acid in a glass bottle, adding purified water to dilute to 1000ml, and mixing uniformly to obtain an enzyme buffer solution;
(3) pepsin: weighing 140 and 141mg of gastric protein powder, pouring the gastric protein powder into a freezing tube, and tightly covering the freezing tube with a cover to obtain the pepsin;
(4) eluent: weighing 175-175.3g of sodium chloride and 88-88.2g of trisodium citrate, adding into a proper container, adding purified water to a constant volume of 1L, stirring until the solution is completely dissolved, and adjusting the pH value to 5.3 +/-0.1 by using 1mol/L hydrochloric acid or 1mol/L sodium hydroxide solution to obtain a 20 XSSC solution;
measuring the obtained 20 XSSC solution into 100ml and 3ml of NP-40, adding into a proper container, adding purified water to a constant volume of 1L, stirring uniformly, and adjusting the pH value to 7.0-7.5 by using 11mol/L hydrochloric acid or 1mol/L sodium hydroxide solution to obtain the eluent.
2. A method of using the pre-hybridization pretreatment reagent according to claim 1, wherein: comprises the steps of reagent preparation, sample treatment and elution after hybridization.
3. The method for using the pre-hybridization pretreatment reagent for a sample according to claim 2, wherein: the reagent preparation comprises the following steps:
s1: pouring 50mL of pretreatment liquid into a dye vat, placing the dye vat in a water bath kettle at 80 +/-1 ℃, and keeping the temperature of the solution in the dye vat at 80 +/-1 ℃ for later use;
s2: pouring 50mL of enzyme buffer solution into a dye vat, placing the dye vat in a water bath kettle at 37 +/-1 ℃, keeping the temperature of the solution in the dye vat at 37 +/-1 ℃ for later use, adding 140mg of pepsin into the enzyme buffer solution when needed, and fully dissolving to obtain a pepsin working solution;
s3: preparing 2 dye vats, adding 50mL of eluent respectively, and placing the dye vats in water bath pots at normal temperature and 73 +/-1 ℃ for later use;
s4: respectively measuring 700mL and 850mL of absolute ethyl alcohol, respectively diluting to 1000mL by using purified water, and sealing and storing at room temperature for later use.
4. The method for using the pre-hybridization pretreatment reagent for a sample according to claim 2, wherein: the sample processing comprises the following steps:
s1 baking slices: slightly drying the slices in the air, and baking the slices on a 65-70 baking sheet machine for 3.8-4h for later use;
s2 dewaxing: at room temperature, immersing the glass slide into dimethylbenzene for dewaxing twice, wherein each time lasts for 10-15 minutes;
s3 dehydration, namely, after dewaxing is finished, soaking the glass slide into absolute ethyl alcohol to dehydrate twice at room temperature for 5 minutes respectively; taking out the slide, and removing residual solution on the slide;
s4 pretreatment: immersing the slide into the pretreatment solution with the temperature of 80 +/-1 ℃ prepared in the first step for treatment for 10 minutes;
s5 removal of solution: taking out the slide, immersing the slide in purified water for 1 minute, and removing residual solution on the slide after taking out;
s6 digestion: placing the glass slide in a pepsin working solution, and digesting for 10-60 minutes at 37 ℃;
s7 soaking: after digestion, the slides were soaked in purified water for 1 minute;
s8 dehydration: after soaking, respectively soaking the glass slides into 70 percent and 85 percent ethanol solutions obtained in the step one and 100 percent ethanol for gradient dehydration for 1 minute respectively;
s9 drying: and (4) taking out the slide after dehydration, and drying at room temperature for later use, thus obtaining the hybridization.
5. The method for using the pre-hybridization pretreatment reagent for a sample according to claim 2, wherein: the post-hybridization elution comprises the following steps:
s1: after the end, taking out the slide from the hybridization instrument, and removing the mounting glue by using tweezers;
s2: at room temperature, immersing the glass slide into the eluent which is arranged in the first step and is reserved at the normal temperature for 5 minutes, and removing the cover glass;
s3: taking out the slide, putting the slide into another eluent which is arranged in the water bath kettle in the step one and is prepared for standby and preheated to 73 +/-1 ℃, and rinsing for 2 minutes;
s4: and taking out the slide, removing residual liquid, and airing in a dark place for later use.
6. The method for using the pretreatment reagent for hybridization of a sample according to claim 3, wherein: the dye vat adopts a Cooprine dye vat.
7. The method for using the pretreatment reagent for hybridization of a sample according to claim 3, wherein: the time at which 140mg of pepsin was added to the enzyme buffer in S2 was before dewaxing the samples.
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| US20210302416A1 (en) * | 2018-07-27 | 2021-09-30 | Veravas, Inc. | Method for detecting biomarkers |
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- 2021-12-10 CN CN202111505159.5A patent/CN114181995A/en active Pending
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102203283A (en) * | 2008-07-21 | 2011-09-28 | 新诊断学股份有限公司 | Methods for the cytological analysis of cervical cells |
| CN105018598A (en) * | 2015-06-05 | 2015-11-04 | 厦门艾德生物医药科技有限公司 | FISH pretreatment method and pretreatment fluid for FFPE sample |
| CN107058593A (en) * | 2017-06-19 | 2017-08-18 | 福建万科药业有限公司 | A kind of hypotype Ph likeALL detection method |
| CN107828862A (en) * | 2017-12-15 | 2018-03-23 | 郑州大学第附属医院 | The preprocess method of lymphoma tissue specimens paraffin embedding slices fluorescence in situ hybridization detection |
| US20210302416A1 (en) * | 2018-07-27 | 2021-09-30 | Veravas, Inc. | Method for detecting biomarkers |
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