WO2019206054A1 - 一种组织芯片量产制作方法 - Google Patents
一种组织芯片量产制作方法 Download PDFInfo
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- WO2019206054A1 WO2019206054A1 PCT/CN2019/083579 CN2019083579W WO2019206054A1 WO 2019206054 A1 WO2019206054 A1 WO 2019206054A1 CN 2019083579 W CN2019083579 W CN 2019083579W WO 2019206054 A1 WO2019206054 A1 WO 2019206054A1
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- 238000004519 manufacturing process Methods 0.000 title claims abstract description 32
- 239000001993 wax Substances 0.000 claims abstract description 42
- 238000000034 method Methods 0.000 claims abstract description 31
- 238000007493 shaping process Methods 0.000 claims abstract description 30
- 239000012188 paraffin wax Substances 0.000 claims abstract description 25
- 239000002245 particle Substances 0.000 claims abstract description 19
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims abstract description 16
- 239000002184 metal Substances 0.000 claims abstract description 13
- 210000004027 cell Anatomy 0.000 claims description 52
- 239000008187 granular material Substances 0.000 claims description 38
- 239000011521 glass Substances 0.000 claims description 20
- 238000010409 ironing Methods 0.000 claims description 19
- 239000003795 chemical substances by application Substances 0.000 claims description 10
- 229920003023 plastic Polymers 0.000 claims description 10
- 210000004748 cultured cell Anatomy 0.000 claims description 8
- 230000008014 freezing Effects 0.000 claims description 8
- 238000007710 freezing Methods 0.000 claims description 8
- 238000010899 nucleation Methods 0.000 claims description 7
- 238000011049 filling Methods 0.000 claims description 5
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 abstract description 2
- 239000002904 solvent Substances 0.000 abstract description 2
- 238000005516 engineering process Methods 0.000 description 13
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Classifications
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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
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
-
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/02—Devices for withdrawing samples
- G01N1/04—Devices for withdrawing samples in the solid state, e.g. by cutting
- G01N1/06—Devices for withdrawing samples in the solid state, e.g. by cutting providing a thin slice, e.g. microtome
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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
- G01N1/36—Embedding or analogous mounting of samples
-
- 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
- G01N1/42—Low-temperature sample treatment, e.g. cryofixation
-
- 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
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
- G01N2001/2866—Grinding or homogeneising
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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
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
- G01N2001/2873—Cutting or cleaving
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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
- G01N1/36—Embedding or analogous mounting of samples
- G01N2001/364—Embedding or analogous mounting of samples using resins, epoxy
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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
- G01N1/36—Embedding or analogous mounting of samples
- G01N2001/368—Mounting multiple samples in one block, e.g. TMA [Tissue Microarrays]
Definitions
- Tissue chip technology is the development and extension of gene chip technology. Like cell chips, protein chips and antibody chips, it is a special biochip technology. Tissue microarray technology can analyze the clinical tissue specimens of dozens or even thousands of different individuals in a pre-designed order on a slide. It is a high-throughput, multi-sample analysis tool. Tissue microarray technology enables researchers to simultaneously analyze hundreds or even thousands of tissue samples simultaneously for the study of one or more specific genes, or expression products associated therewith. The technology can be combined with DNA, RNA, protein, antibody and other technologies, combined with traditional pathology, histochemistry and immunohistochemistry, at the three levels of gene, gene transcription and biological functions of related expression products. the study.
- the tissue of the tissue mixer after mixing has an irregular shape, but still retains a tissue structure.
- the tissue particles have a size of from 0.01 cm to 0.3 cm.
- the plastic tube is placed in the groove on the ironing table of the tissue chip embedding machine; the tissue particles are stuffed into the shaping tube, and the melted paraffin is added to fill the gap between the tissue grains in the shaping tube; the tissue particles and paraffin are filled
- the shaping tube filling the gap is transferred from the ironing table to the freezing station of the tissue chip embedding machine to obtain a shaped tube with the shaped tissue core, which is stored in numbers, and the shaped tube is stripped before use to obtain shaping. After the tissue core.
- the method for adhering the shaped tissue or cells to the metal plate comprises: placing the metal plate in a refrigerator, marking the metal plate with a positioning point, and taking out the frozen shaped tissue or cells, Record the position on the positioning plate, apply a little embedding agent to one end of the shaped tissue or cell, and stick it one by one to the positioning point of the metal plate, and then fill the shaped tissue or interstitial space with the embedding agent.
- the embedding agent is frozen with the shaped tissue or cells, and sliced with a cryostat to obtain a tissue or a cell chip.
- Step 5 After the seeding is completed, the acceptor wax mold planting surface is directed toward the glass plate, and placed in a constant temperature oven at 56 ° C for 60 minutes. After polymerization, the acceptor wax mold and the glass plate are simultaneously placed in a refrigerator at 4 ° C. The glass plate is withdrawn, that is, an acceptor wax pattern of the inoculated tissue or cells is obtained, and the tissue chip is obtained by slicing in the direction parallel to the glass plate.
- the five tissue cores of the same case can participate in the preparation of five tissue chips. Thus, 10,000 chips can be obtained.
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- General Health & Medical Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Sampling And Sample Adjustment (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
Claims (10)
- 一种组织芯片量产制作方法,其特征在于,包括:将组织粒、培养的细胞或收集的脱落细胞用塑形装置塑形,所述的组织粒通过采用组织搅拌机搅碎新鲜组织、福尔马林或其他溶剂固定的组织、或石蜡包埋的组织块,将塑形后的组织或细胞种入受体蜡模,切片,即获得组织芯片。
- 如权利要求1所述的组织芯片量产制作方法,其特征在于,所述的组织搅拌机搅拌后的组织粒形状不规则,但仍保留有组织结构。
- 如权利要求1所述的组织芯片量产制作方法,其特征在于,所述的组织粒大小在0.01cm~0.3cm。
- 如权利要求1所述的组织芯片量产制作方法,其特征在于,采用组织搅拌机搅碎福尔马林或其他溶剂固定的组织形成的组织粒的塑形方法包括:收集采用组织搅拌机搅碎福尔马林或其他溶剂固定的组织形成的组织粒,用棉布包裹,编号后脱水浸蜡;打开组织芯片包埋机,将组织粒放在组织芯片包埋机的烫台上,在50℃~70℃将其上的石蜡融化,即为待塑形的组织粒;将塑形管置入组织芯片包埋机烫台上的凹槽中;将组织粒塞入塑形管内,加入融化的石蜡填充塑形管内组织粒间的缝隙;将充满组织粒和石蜡填充缝隙的塑形管从烫台转入组织芯片包埋机的冻台聚合,获得装有塑形后的组织芯的塑形管,编号存放,使用前将塑形管剥除,得到塑形后的组织芯。
- 如权利要求1所述的组织芯片量产制作方法,其特征在于,所述的将塑形后的组织或细胞种入受体蜡模的方法包括:将受体蜡模放在玻璃板上,置入50℃-70℃恒温烤箱20-40分钟,使受体蜡模变软,在定位板或定位纸上记录相应位置的组织或细胞编号,用引导针在受体蜡模相应位置钻孔,将塑形后的组织或细胞逐个种入,种入完成后,将种入面朝向玻璃板,放在50℃-70℃恒温烤箱聚合1-3小时,将受体蜡模和玻璃板同时置入2-6℃冰箱,将玻璃板撤出,即获得接种组织或细胞的受体蜡模,沿平行于玻璃板方向切片即获得组织芯片。
- 如权利要求1所述的组织芯片量产制作方法,其特征在于,所述的采用组织搅拌机搅碎石蜡包埋的组织块形成的组织粒的塑形方法包括:将组织搅拌机搅碎石蜡包埋的组织块形成的组织粒收集起来,将塑形管置入组织芯片包埋机烫台上的凹槽中;将组织粒塞入塑形管内,加入融化的石蜡填充塑形管内组织粒间的缝隙;将充满组织粒和石蜡填充缝隙的塑形管从烫台转入组织芯片包埋机的冻台聚合,获得装有塑形后的组织芯的塑形管,编号存放,使用前将塑形管剥除,得到塑形后的组织芯。
- 如权利要求1所述的组织芯片量产制作方法,其特征在于,所述的采用组织搅拌机搅碎新鲜组织形成的组织粒的塑形方法包括:将组织粒用吸管移入至新鲜组织粒塑形板上,新鲜组织粒塑形板为有弹性的塑料板,塑料板上有凹槽,将新鲜组织粒吸入凹槽,将新鲜组织粒塑形板置于-15~-25℃冰箱,即形成新鲜组织冻存柱,编号, 备用。
- 如权利要求1所述的组织芯片量产制作方法,其特征在于,所述的将塑形后的组织或细胞粘连在金属板上的方法包括:将金属板置于冰箱中,金属板上标记有定位点,取出塑形后的组织或细胞,在定位板上记录位置,将塑形后的组织或细胞的一端涂抹少许包埋剂,逐一粘连在金属板的定位点上,之后用包埋剂将塑形后的组织或细胞间的空隙填充,包埋剂与塑形后的组织或细胞冷冻为一体,用冷冻切片机切片,即获得组织或细胞芯片。
- 如权利要求1所述的组织芯片量产制作方法,其特征在于,所述的将培养的细胞或收集的脱落细胞用塑形装置塑形的方法包括:将培养的细胞或收集的脱落细胞直接采用常规方法固定、脱水、浸蜡,将塑形管置入组织芯片包埋机烫台上的凹槽中,将浸蜡后的细胞置入塑形管内,加入融化的石蜡填充缝隙;将塑形管从烫台转入组织芯片包埋机的冻台聚合,获得塑形后的细胞,编号存放。
- 如权利要求4、6或9所述的组织芯片量产制作方法,其特征在于,所述的塑形管为1~2cm长,直径0.1~0.3cm的圆柱形塑料管,或者,所述的塑形管为长为1~2cm,宽为0.1~0.5cm,高为0.1~0.5cm的长方体形塑形管。
Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
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MYPI2020005582A MY196704A (en) | 2018-04-25 | 2019-04-20 | Mass Production Manufacturing Method For Tissue Chip |
KR1020207031934A KR102458364B1 (ko) | 2018-04-25 | 2019-04-20 | 조직 칩 양산 제작 방법 |
JP2020560136A JP7111833B2 (ja) | 2018-04-25 | 2019-04-20 | 組織チップの量産製造方法 |
US17/078,372 US20210041333A1 (en) | 2018-04-25 | 2020-10-23 | Mass production manufacturing method for tissue chip |
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CN201810381966.2A CN108593380B (zh) | 2018-04-25 | 2018-04-25 | 一种组织芯片量产制作方法 |
CN201810381966.2 | 2018-04-25 |
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US17/078,372 Continuation US20210041333A1 (en) | 2018-04-25 | 2020-10-23 | Mass production manufacturing method for tissue chip |
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US (1) | US20210041333A1 (zh) |
JP (1) | JP7111833B2 (zh) |
KR (1) | KR102458364B1 (zh) |
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Cited By (1)
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CN111088216A (zh) * | 2019-12-16 | 2020-05-01 | 齐妍 | 一种用于石蜡包埋组织分离检测细胞的方法 |
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ES2920502T3 (es) * | 2017-12-06 | 2022-08-04 | Genentech Inc | Controles sintéticos para inmunohistoquímica |
CN108593380B (zh) * | 2018-04-25 | 2021-05-18 | 复旦大学附属中山医院 | 一种组织芯片量产制作方法 |
CN113759143B (zh) * | 2021-09-01 | 2024-04-26 | 广州耐确医疗器械有限责任公司 | 一种新型全自动组织芯片仪 |
CN114002030A (zh) * | 2021-10-15 | 2022-02-01 | 北京龙迈达斯科技开发有限公司 | 一种复合型组织芯片及其制备方法 |
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CN101694429A (zh) * | 2009-10-15 | 2010-04-14 | 复旦大学附属中山医院 | 一种石蜡组织芯片包埋方法 |
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CN100451649C (zh) * | 2004-01-08 | 2009-01-14 | 中南大学湘雅医学院肿瘤研究所 | 一种细胞芯片制作方法及其器具 |
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CN108593380A (zh) | 2018-09-28 |
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