CN103900864A - Exfoliated cell chip - Google Patents
Exfoliated cell chip Download PDFInfo
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- CN103900864A CN103900864A CN201210570306.1A CN201210570306A CN103900864A CN 103900864 A CN103900864 A CN 103900864A CN 201210570306 A CN201210570306 A CN 201210570306A CN 103900864 A CN103900864 A CN 103900864A
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Abstract
The present invention provides a technology and a method for preparing a high-throughput exfoliated cell chip. According to the present invention, a cervix brush is adopted to collect cervix exfoliated cells of 26 cervical lesions patients and are immediately placed into a cell preservation solution, centrifugation is performed, the supernatant is removed, a cell immobilization solution is added to immobilize cell precipitate, dehydration transparency and dipping in wax are performed, a tissue chip instrument is adopted to prepare an exfoliated cell paraffin chip and an exfoliated cell paraffin block array, and paraffin slicing is performed to prepare the exfoliated cell chip; and the defect of the conventional cytology detection is overcome, and the prepared exfoliated cell chip can be used for pathological diagnosis, IHC, in situ hybridization and other molecular medicine researches.
Description
Technical field
The invention belongs to biochip field, relate to a kind of for the preparation of microarray come off biochip and the preparation method of chip.
Background technology
Biochip refers to the high flux microarray of the high density tissue, cell, protein, nucleic acid, carbohydrate and the other biological component that are coated on the solid support such as silicon chip, nylon membrane.Utilize high flux biochip, hybridize and staining technique by specific molecular, examined samples are detected, can be accurately, a large amount of Molecular Detection information of quick obtaining.The fields such as life science, clinical medicine molecular diagnosis and the examination of biological antibody targeted drug and checking are widely used at present.
Exfoliative cytology inspection is one of important means of early diagnosis cancer, and its fundamental procedure is: centrifugal collection cast-off cells, by cell precipitation slowly drip on microslide, prepare cell smear, sheet pathological diagnosis is read in dyeing.Its deficiency is: 1. the as easy as rolling off a log phenomenon that occurs cell overlap on prepared cell smear, causes tumour cell almost illegible; 2. tumour cell is likely organized or organizes agglomerate to cover by other fragments, causes testing result false negative; 3. Exfoliated tumor cells agglomerate can not effectively disperse, and causes pathological diagnosis somatotype difficulty; 4. in dyeing course,, by the effectively tumour cell of absorption of microslide, likely do not washed away, caused false negative result; 5. owing to being the cytology dyeing adopting for cell, be difficult to tumour cell to carry out clear and definite pathological staging and somatotype.Prepared one or several cell smears for pathological staining diagnosis after, there is no cast-off cells sample for further detecting and other research experiments.
Summary of the invention
The present invention aims to provide a kind of technology and method of preparing high flux cast-off cells chip.Overcome the defect that above-mentioned exfoliative cytology checks, prepared cast-off cells chip can be used for pathological diagnosis, IHC, the medical research of in situ hybridization equimolecular.
The present invention is achieved through the following technical solutions goal of the invention.Utilize Uterine neck bush to collect 26 routine cervical lesions patients' cervical exfoliated cell, put into immediately cell-preservation liquid; Centrifugal, abandon supernatant, add cell immobile liquid fixed cell precipitation; Transparent, waxdip dewaters.Utilize organization chip instrument to prepare cast-off cells paraffin core, cast-off cells paraffin mass array, paraffin section is prepared cast-off cells chip continuously.Prepared cast-off cells chip pathological staining effect is significantly better than conventional cell smear Color.Due to cast-off cells highly dense, paraffin section thinner (4um), and can serial section, the cell precipitation piece of about 1.7mm3 can be made 250 of sections in theory, has greatly improved the service efficiency of cast-off cells.
Accompanying drawing explanation
Fig. 1 is the schematic diagram of the hollow paraffin mass wax-pattern of array: all apertures are 1.5mm, and pitch of holes is 2mm.
Fig. 2 is the schematic diagram of cast-off cells array paraffin mass: cast-off cells paraffin stem stem diameter is 1.5mm, and cast-off cells paraffin stem stem spacing is 2mm.
Fig. 3 is the schematic diagram of cast-off cells chip: cast-off cells paraffin chip spot diameter is 1.5mm, and cast-off cells paraffin chip dot spacing is 2mm.Mark 1: be the frosted marked region on cast-off cells chip solid phase carrier (adhesion microslide).
Embodiment
Embodiment: on the basis of fully informed consent, collect 42 routine cervical lesions patients' cervical exfoliated cell, prepare cast-off cells chip.
1.1 cast-off cells are collected: utilize Uterine neck bush to collect 42 routine cervical lesions patients' cervical exfoliated cell, put into immediately the cell on the abundant wash-out Uterine neck bush of cell-preservation liquid.Centrifugal, remove supernatant, collect cast-off cells precipitation.
1.2 cast-off cells are fixed: in cast-off cells precipitation, add 1ml cell immobile liquid, fully mix, room temperature is fixed 1 hour.
1.3 dehydration embeddings: centrifugal collection cast-off cells, neutral PBS damping fluid washing cast-off cells 2 times.After the dehydration of alcohol gradient, degreasing is transparent, the cast-off cells that centrifugal collection and treatment is good.Cast-off cells are added in the paraffin of 50ul preheating thawing, mix.The cooling cast-off cells paraffin mass that is fixed into.
1.4 cast-off cells wax cores: utilize organization chip preparing instrument paraffin core sampler, drilling through diameter is the cast-off cells wax core of 1.5mm.
1.5 acceptor wax-patterns: prepare blank paraffin mass, utilize organization chip preparing instrument to locate and drill through aperture the hollow acceptor wax-pattern that is 1.5ml, as accompanying drawing 1.Pitch of holes is 2mm.
1.6 cast-off cells arrays: according to the design of cast-off cells chip, 26 routine sperm paraffin cores are placed to corresponding blank well site in hollow acceptor wax-pattern one by one altogether, flush as benchmark, as accompanying drawing 2 take array surface.
1.7 arrays merge: sperm array is placed in to drying box, regulates drying box temperature and time, the distortion of guaranteeing that all cast-off cells paraffin cores and acceptor wax-pattern fully merge and cast-off cells paraffin core is not shifted.
1.8 paraffin sections: utilize paraffin slicing machine section serial section, the thick 4um of sheet.By on paraffin section tiling and adhesion microslide, as accompanying drawing 3.Perform mark at microslide marked region.
1.9 cast-off cells chip manufacturings are complete.Prepared cast-off cells chip can be used for pathological diagnosis and other molecular medicine experimental studies such as conventional H E dyeing, SABC, hybridization histochemistry.
Claims (4)
1. a preparation method for cast-off cells chip, is characterized in that: comprise the following steps:
The first step, cast-off cells are collected: utilize Uterine neck bush to collect 42 routine cervical lesions patients' cervical exfoliated cell, put into immediately the cell on the abundant wash-out Uterine neck bush of cell-preservation liquid.Centrifugal, remove supernatant, collect cast-off cells precipitation.
Second step, cast-off cells are fixed: in cast-off cells precipitation, add 1ml cell immobile liquid, fully mix, room temperature is fixed 1 hour.
The 3rd step, dehydration embedding: centrifugal collection cast-off cells, neutral PBS damping fluid washing cast-off cells 2 times.After the dehydration of alcohol gradient, degreasing is transparent, the cast-off cells that centrifugal collection and treatment is good.Cast-off cells are added in the paraffin of 50ul preheating thawing, mix.The cooling cast-off cells paraffin mass that is fixed into.
The 4th step, cast-off cells wax core: utilize organization chip preparing instrument paraffin core sampler, drilling through diameter is the cast-off cells wax core of 1.5mm.
The 5th step, acceptor wax-pattern: prepare blank paraffin mass, utilize organization chip preparing instrument to locate and drill through aperture the hollow acceptor wax-pattern that is 1.5ml, as accompanying drawing 1.Pitch of holes is 2mm.
The 6th step, cast-off cells array: according to the design of cast-off cells chip, 26 routine sperm paraffin cores are placed to corresponding blank well site in hollow acceptor wax-pattern one by one altogether, flush as benchmark, as accompanying drawing 2 take array surface.
The 7th step, array merges: sperm array is placed in to drying box, regulates drying box temperature and time, the distortion of guaranteeing that all cast-off cells paraffin cores and acceptor wax-pattern fully merge and cast-off cells paraffin core is not shifted.
The 8th step, paraffin section: utilize paraffin slicing machine microsection manufacture paraffin section, the thick 4um of sheet.By on paraffin section tiling and adhesion microslide, as accompanying drawing 3.Perform mark at microslide marked region.
2. the preparation method of a kind of cast-off cells chip according to claim 1, is characterized in that: cast-off cells refer to the cast-off cells in various Celom liquid loadings such as comprising cervical exfoliated cell, tracheae/bronchus cast-off cells, vaginal exfoliated and ascites/hydrothorax.
3. the preparation method of a kind of cast-off cells chip according to claim 1, is characterized in that: cast-off cells, after dehydration embedding treatment, are directly used in the preparation of cast-off cells paraffin core.
4. the preparation method of a kind of cast-off cells chip according to claim 1, it is characterized in that: cast-off cells chip is rectangular array composition, sperm paraffin chip spot diameter is (but being not limited to) 1.5mm, and cast-off cells paraffin chip dot spacing is (but being not limited to) 2mm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201210570306.1A CN103900864A (en) | 2012-12-25 | 2012-12-25 | Exfoliated cell chip |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201210570306.1A CN103900864A (en) | 2012-12-25 | 2012-12-25 | Exfoliated cell chip |
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| CN103900864A true CN103900864A (en) | 2014-07-02 |
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| CN201210570306.1A Pending CN103900864A (en) | 2012-12-25 | 2012-12-25 | Exfoliated cell chip |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104569397A (en) * | 2015-01-30 | 2015-04-29 | 卫生部北京医院 | Quality control sample for detecting breast cancer and preparation method of quality control sample |
| CN104745670A (en) * | 2015-04-16 | 2015-07-01 | 黄小军 | Preparation method of cell chip and application of cell chip in tumour screening field |
| CN105241726A (en) * | 2015-09-14 | 2016-01-13 | 黄小军 | Immunohistochemical staining method and applications of immunohistochemical staining method in cervical tumorous lesion screening |
| CN107192594A (en) * | 2017-07-21 | 2017-09-22 | 上海市第妇婴保健院 | A kind of decidua and fine hair pairing organization chip, preparation method and applications |
| CN108593380A (en) * | 2018-04-25 | 2018-09-28 | 复旦大学附属中山医院 | A kind of organization chip volume production production method |
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| CN1356553A (en) * | 2001-09-13 | 2002-07-03 | 陕西超英生物医学研究开发有限公司 | Tissue microarray biochip |
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| CN101231282A (en) * | 2007-01-23 | 2008-07-30 | 北京市农林科学院 | Organization chip for researching functional genome as well as preparation method and application thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104569397A (en) * | 2015-01-30 | 2015-04-29 | 卫生部北京医院 | Quality control sample for detecting breast cancer and preparation method of quality control sample |
| CN104745670A (en) * | 2015-04-16 | 2015-07-01 | 黄小军 | Preparation method of cell chip and application of cell chip in tumour screening field |
| CN105241726A (en) * | 2015-09-14 | 2016-01-13 | 黄小军 | Immunohistochemical staining method and applications of immunohistochemical staining method in cervical tumorous lesion screening |
| CN107192594A (en) * | 2017-07-21 | 2017-09-22 | 上海市第妇婴保健院 | A kind of decidua and fine hair pairing organization chip, preparation method and applications |
| CN107192594B (en) * | 2017-07-21 | 2019-11-05 | 上海市第一妇婴保健院 | A kind of decidua and villus match organization chip, preparation method and applications |
| CN108593380A (en) * | 2018-04-25 | 2018-09-28 | 复旦大学附属中山医院 | A kind of organization chip volume production production method |
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Application publication date: 20140702 |