CN110082180A - A method of detection sample is prepared based on fresh cancerous tissue paster method - Google Patents
A method of detection sample is prepared based on fresh cancerous tissue paster method Download PDFInfo
- Publication number
- CN110082180A CN110082180A CN201910378420.6A CN201910378420A CN110082180A CN 110082180 A CN110082180 A CN 110082180A CN 201910378420 A CN201910378420 A CN 201910378420A CN 110082180 A CN110082180 A CN 110082180A
- Authority
- CN
- China
- Prior art keywords
- cancer
- sample
- tissue
- flesh tissue
- plane support
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- 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
- G01N1/2813—Producing thin layers of samples on a substrate, e.g. smearing, spinning-on
-
- 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"
-
- 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
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5091—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing the pathological state of an organism
-
- 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"
- G01N2021/6439—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" with indicators, stains, dyes, tags, labels, marks
Landscapes
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Physics & Mathematics (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Pathology (AREA)
- Analytical Chemistry (AREA)
- Urology & Nephrology (AREA)
- Molecular Biology (AREA)
- Hematology (AREA)
- Biomedical Technology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Physiology (AREA)
- Microbiology (AREA)
- Optics & Photonics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of methods for preparing detection sample based on fresh cancerous tissue paster method, belong to field of biomedicine.Method includes the following steps: (1) obtains cancer flesh tissue；(2) cell of cancer flesh tissue is affixed on plane support；(3) plane support is placed in drying at room temperature 24-30h to get cancer flesh tissue sample is arrived.Using method of the invention, cancer flesh tissue sample can be quickly prepared, it is simple, time saving, laborsaving；The cancer flesh tissue sample of preparation is detected for FISH, while time saving, laborsaving, province takes, and testing result is clear, accurately.
Description
Technical field
The invention belongs to fields of biomedicine, and in particular, to one kind is based on fresh cancerous tissue paster method preparation detection mark
This method.
Background technique
Fluorescence in situ hybridization technique (FISH) refers to that using the DNA single-chain nucleic acid of specific process fluorescent marker be probe, presses
Single-chain nucleic acid in base complementrity principle and material to be checked is specifically bound into double-strandednucleic acid, detects chromosome specific cdna number
Molecular pathology technology that is changing and detecting the exceptions such as gene magnification, missing or Diagnosis of malignant cell.It is widely used at present
Malignant cell detection, genetic disease diagnosis, pre-natal diagnosis and malignant tumor patient individuation targeted therapy before base
Gene-amplification detection etc..Compared with traditional pathological diagnosis, the sensibility and specificity of FISH technology is significantly high.
It needs to make sample with FISH technology detection gene.Traditional production sample method mainly uses paraffin-embedded tissue
Sample.The sample preparation method mainly comprises the steps that (1) formaldehyde is fixed；(2) paraffin embedding；(3) it is sliced；(4) before detecting
Carry out a series of tedious steps such as roasting piece, dewaxing, pretreatment, digestion.
Summary of the invention
In order to solve the above-mentioned technical problem, inventor has carried out a large amount of explorations, finds a kind of new cancer flesh tissue mark
This preparation method has unexpected technical effect, thereby completing the present invention.
The present invention provides a kind of methods for preparing detection sample based on fresh cancerous tissue paster method, include the following steps
(1) cancer flesh tissue is obtained；
(2) cell of cancer flesh tissue is affixed on plane support；
(3) plane support is placed in drying at room temperature 24-30h to get cancer flesh tissue sample is arrived.
In the present invention, the cancer is not particularly limited, and can be any type of solid type tumour.
In some embodiments of the present invention, the cancer is selected from the cancer of the esophagus, lung cancer, cardia cancer, liver cancer.In the present invention
A preferred embodiment in, the cancer be the cancer of the esophagus.
In some embodiments of the present invention, the plane support is glass slide.
In some embodiments of the present invention, the room temperature refers to 15 DEG C~30 DEG C, such as 15,20,25 or 30 DEG C.?
In one embodiment of the invention, the room temperature refers to 25 DEG C.
In some embodiments of the present invention, step (3) by plane support be placed in drying at room temperature 24,25,27,27,
28,29 or 30h.In some specific embodiments of the invention, plane support is placed in drying at room temperature for 24 hours by step (3).
In the present invention, the sample is detected for FISH.
In some embodiments of the present invention, sample low temperature environment is placed in front of carrying out FISH detection to save.
In some specific embodiments of the invention, the low temperature refers to -20 DEG C~-80 DEG C.
In one embodiment of the invention, the low temperature refers to -20 DEG C.
In another specific embodiment of the invention, the low temperature refers to -80 DEG C.
In the present invention, it takes a step forward in progress FISH detection and includes the steps that for sample being placed in 45 DEG C of dry 5min.
Beneficial effects of the present invention
Method of the invention has the advantages that compared with the existing technology
(1) utilize method of the invention, can quickly prepare cancer flesh tissue sample, greatly reduce formaldehyde fix, stone
Consumption in time and expense caused by the technologies such as wax embedding, slice, it is simple, time saving, laborsaving；
(2) the cancer flesh tissue sample prepared using method of the invention, further reducing roasting piece before detection (needs
Overnight), the step of dewaxing (about 40min), pretreatment (about 20min) and digestion (15min), so that detection is more time saving, laborsaving, saves
Take.
(3) FISH detection is carried out using cancer flesh tissue sample prepared by method of the invention, testing result is clear, quasi-
Really.
Detailed description of the invention
Fig. 1 shows the sample (left side) of the method for the present invention preparation and the sample (right side) using paraffin embedding techniques preparation.
Fig. 2 shows the testing results of the detection sample of preparation.A: normal cell core-FISH image；B: esophageal cancer cell
Core-FISH image；C: normal tissue-IHC image: D: the cancer of the esophagus-IHC image.
Specific embodiment
In order to which the technical problems, technical solutions and beneficial effects solved by the present invention is more clearly understood, below in conjunction with
Embodiment, the present invention will be described in further detail.
Embodiment
Following example is used herein to demonstration the preferred embodiments of the invention.Those skilled in the art, it will be appreciated that under
State the technology disclosed in example represent inventor discovery can be used for implementing technology of the invention, therefore can be considered as implementation this
The preferred embodiment of invention.But those skilled in the art should be understood that specific reality disclosed herein according to this specification
Many modifications can be made by applying example, still can be obtained identical or similar as a result, rather than away from the spirit or scope of the present invention.
Unless otherwise defined, the term of all technologies as used herein and science, and the technology in fields of the present invention
Personnel institute is normally understood equivalent in meaning, and being disclosed reference and their materials of reference will all be incorporated.
Those skilled in the art will recognize or just will appreciate that by routine test many described here
Invention specific embodiment many equivalent technologies.These will equally be comprised in claims.
Embodiment
The present embodiment is utilized respectively flesh tissue paster method and paraffin imbedding preparation cancerous tissue sample.
Cancerous tissue used in the present embodiment is the cancer of the esophagus flesh tissue cut off in art.
1. paraffin imbedding prepares tissue specimen
It is fixed: tissue being placed in 4% formalin and is fixed for 24 hours.
It rinses: tissue being placed in fixed window and rinses 5min with flowing water.
Dehydration: 75% ethyl alcohol 30 minutes, 85% ethyl alcohol 30 minutes, 95% ethyl alcohol 40min, dehydrated alcohol I30min are anhydrous
Ethyl alcohol II30min.
It is transparent: tissue after dehydration is directly immersed in dimethylbenzene clarifier (dimethylbenzene I, dimethylbenzene II), it is transparent every time
10min。
Waxdip: first immersing soft wax I1h or so for tissue, then immerse soft wax II1h, hard wax I40min, hard wax II1h.
Embedding: by the hard tool of wax liquor injection embedding, tissue block is laid flat in wax liquor rapidly, ajusts and paves, then move
To cooling bench, it is freezing together tissue block with wax liquor.
Slice, bonding die and roasting piece: slice thickness is 5 μm, is invested in wax disk(-sc) jail on glass slide with bonding die agent, it is inhaled with filter paper
It takes extra moisture, then glass slide is put in 60 DEG C of exhibition piece platforms and dries (overnight), that is, prepare tissue specimen (such as Fig. 1 is right).
2. flesh tissue paster method prepares tissue specimen
The cell print of cancer of the esophagus flesh tissue is affixed on glass slide, glass slide containing cell is then placed in room temperature
(25 DEG C) dryings for 24 hours, obtain tissue specimen (such as Fig. 1 is left), are placed in -80 DEG C of refrigerators and save backup.
3.FISH detection
The tissue specimen respectively prepared by above two method carries out FISH detection.
Using tissue specimen prepared by paraffin embedding techniques carried out before being detected roasting piece (overnight), dewax (40min),
Pre-process (20min) and digestion (15min).
It is directly detected after 45 DEG C of dry 5min using tissue specimen prepared by flesh tissue paster method.
As a result, it has been found that (as shown in Figure 2): the tissue specimen prepared using flesh tissue paster method, the neoplastic cell nuclei of detection
Middle signal is strong and bright, is significantly better than the sample prepared using paraffin embedding techniques.
All references mentioned in the present invention is incorporated herein by reference, independent just as each document
It is incorporated as with reference to such.In addition, it should also be understood that, after reading the above teachings of the present invention, those skilled in the art can
To make various changes or modifications to the present invention, such equivalent forms equally fall within model defined by the application the appended claims
It encloses.
Claims (10)
1. a kind of method for preparing detection sample based on fresh cancerous tissue paster method, which comprises the following steps:
(1) cancer flesh tissue is obtained；
(2) cell of cancer flesh tissue is affixed on plane support；
(3) plane support is placed in drying at room temperature 24-30h to get cancer flesh tissue sample is arrived.
2. the method according to claim 1, wherein the cancer is selected from the cancer of the esophagus, lung cancer, cardia cancer, liver cancer.
3. the method according to claim 1, wherein the plane support is glass slide.
4. the method according to claim 1, wherein the room temperature refers to 15 DEG C~30 DEG C.
5. according to the method described in claim 4, it is characterized in that, the room temperature refers to 25 DEG C.
6. according to the method described in claim 5, it is characterized in that, plane support is placed in drying at room temperature 24 hours.
7. the method according to claim 1, wherein the sample is detected for FISH.
8. being saved the method according to the description of claim 7 is characterized in that sample is placed in low temperature environment before detection.
9. according to the method described in claim 8, it is characterized in that, the low temperature refers to -20 DEG C~-80 DEG C.
10. method according to claim 8 or claim 9, which is characterized in that before detection further comprise that sample is placed in 45 DEG C
The step of dry 5min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910378420.6A CN110082180A (en) | 2019-05-08 | 2019-05-08 | A method of detection sample is prepared based on fresh cancerous tissue paster method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910378420.6A CN110082180A (en) | 2019-05-08 | 2019-05-08 | A method of detection sample is prepared based on fresh cancerous tissue paster method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110082180A true CN110082180A (en) | 2019-08-02 |
Family
ID=67419112
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910378420.6A Pending CN110082180A (en) | 2019-05-08 | 2019-05-08 | A method of detection sample is prepared based on fresh cancerous tissue paster method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110082180A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108663251A (en) * | 2018-05-16 | 2018-10-16 | 绍兴文理学院 | A kind of tumor lympha closes the processing method for sweeping flesh tissue sample |
| CN108956242A (en) * | 2018-05-16 | 2018-12-07 | 绍兴文理学院 | A kind of fixing process method of mammary cancer armpit lymph gland in vitro tissue sample |
-
2019
- 2019-05-08 CN CN201910378420.6A patent/CN110082180A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108663251A (en) * | 2018-05-16 | 2018-10-16 | 绍兴文理学院 | A kind of tumor lympha closes the processing method for sweeping flesh tissue sample |
| CN108956242A (en) * | 2018-05-16 | 2018-12-07 | 绍兴文理学院 | A kind of fixing process method of mammary cancer armpit lymph gland in vitro tissue sample |
Non-Patent Citations (5)
| Title |
|---|
| 余永强 等: "《现代医院诊疗常规》", 30 September 2012, 安徽科学技术出版社 * |
| 徐肇玥 等: "《病毒性肝炎研究进展》", 30 April 1980, 上海科学技术出版社 * |
| 杨荷英 等: "《实用临床医学检验》", 30 June 2018, 上海交通大学出版社 * |
| 武汉医学院第一附属医院: "《临床细胞学图谱》", 31 October 1972, 武汉人民出版社 * |
| 王燕蓉 等: "《形态学实用技术》", 30 April 2010, 第二军医大学出版社 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US11970739B2 (en) | Multiplex capture of gene and protein expression from a biological sample | |
| US11519908B2 (en) | Matrix for receiving tissue samples | |
| Maniatis et al. | Spatially resolved transcriptomics and its applications in cancer | |
| CN102238991B (en) | The biomolecule that parallel extracting is different from the tissue that formalin is fixed | |
| Chetcuti et al. | Identification of differentially expressed genes in organ‐confined prostate cancer by gene expression array | |
| EP2239555A1 (en) | Novel method of preparing specimen having excellent abilities to maintain tissue morphology and maintain nucleic acid qualities | |
| CN101988061A (en) | Breast cancer detecting marker as well as detecting method, kit and biological chip thereof | |
| CN105950539B (en) | Construction method and application of P-glycoprotein model for human small intestine 3D organ research | |
| CN107574168A (en) | A kind of miRNA of regulation and control red tilapia body-color diversity and its application | |
| Takikita et al. | Tissue microarrays enabling high-throughput molecular pathology | |
| Skubitz et al. | Differential gene expression identifies subgroups of renal cell carcinoma | |
| CN105925719A (en) | Gene related to liver cancer differentiation and application of gene | |
| Chen et al. | Advances in cancer tissue microarray technology: Towards improved understanding and diagnostics | |
| CN110082180A (en) | A method of detection sample is prepared based on fresh cancerous tissue paster method | |
| CN110273001A (en) | One kind circRNA relevant to prostate cancer and its application | |
| Kosuge et al. | Nuclear features of infiltrating urothelial carcinoma are distinguished from low-grade noninvasive papillary urothelial carcinoma by image analysis | |
| Duderstadt et al. | A method to pre-screen rat mammary gland whole-mounts prior to RNAscope | |
| CN108588222B (en) | A kind of hepatocellular carcinoma auxiliary diagnosis or disease surveillance kit and application | |
| CN116773790B (en) | Preparation method and application of tumor tissue HER2 gradient detection product | |
| CN109628594A (en) | One kind long-chain non-coding RNA relevant to liver cancer and its application | |
| CN106460072A (en) | Spatial molecular profiling of solid biological masses and profile storage | |
| CN118086511B (en) | Stomach mucosa intestinal metaplasia subtype biomarker FOLH1 and application thereof | |
| CA3058904A1 (en) | High precision micropurification system and methods of use | |
| Block et al. | P-Glucuronidase enzyme histochemistry on semithin sections of plastic-embedded Arabidopsis explants | |
| Chung et al. | Tissue microarrays in biomedical research |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20190802 |