KR100386340B1 - Multiple vaccuum shield of hybridization cover seal - Google Patents
Multiple vaccuum shield of hybridization cover seal Download PDFInfo
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- KR100386340B1 KR100386340B1 KR10-2001-0000940A KR20010000940A KR100386340B1 KR 100386340 B1 KR100386340 B1 KR 100386340B1 KR 20010000940 A KR20010000940 A KR 20010000940A KR 100386340 B1 KR100386340 B1 KR 100386340B1
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- cover seal
- diaphragm
- diaphragms
- hybridization
- nucleic acid
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- 238000009396 hybridization Methods 0.000 title claims abstract description 19
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 27
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 27
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 27
- 238000006243 chemical reaction Methods 0.000 claims abstract description 25
- 238000001179 sorption measurement Methods 0.000 claims abstract description 19
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 14
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 14
- 238000002493 microarray Methods 0.000 claims abstract description 5
- 229920001296 polysiloxane Polymers 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 6
- 239000007850 fluorescent dye Substances 0.000 claims 1
- 239000011521 glass Substances 0.000 abstract description 15
- 241000283216 Phocidae Species 0.000 description 30
- 239000000243 solution Substances 0.000 description 11
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 8
- 239000000853 adhesive Substances 0.000 description 8
- 230000001070 adhesive effect Effects 0.000 description 8
- 229910052710 silicon Inorganic materials 0.000 description 8
- 239000010703 silicon Substances 0.000 description 8
- 239000007853 buffer solution Substances 0.000 description 5
- 238000011109 contamination Methods 0.000 description 5
- 238000012757 fluorescence staining Methods 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 4
- 239000000872 buffer Substances 0.000 description 3
- 230000008020 evaporation Effects 0.000 description 2
- 238000001704 evaporation Methods 0.000 description 2
- 230000014509 gene expression Effects 0.000 description 2
- 238000012744 immunostaining Methods 0.000 description 2
- 238000011065 in-situ storage Methods 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 102000004859 Cholecystokinin Receptors Human genes 0.000 description 1
- 108090001085 Cholecystokinin Receptors Proteins 0.000 description 1
- 102000003688 G-Protein-Coupled Receptors Human genes 0.000 description 1
- 108090000045 G-Protein-Coupled Receptors Proteins 0.000 description 1
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 1
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 1
- 108091005461 Nucleic proteins Proteins 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000003139 buffering effect Effects 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000006059 cover glass Substances 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000012202 endocytosis Effects 0.000 description 1
- 239000005090 green fluorescent protein Substances 0.000 description 1
- 238000010191 image analysis Methods 0.000 description 1
- 238000011532 immunohistochemical staining Methods 0.000 description 1
- 238000007901 in situ hybridization Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 208000003154 papilloma Diseases 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 201000010159 squamous cell papilloma Diseases 0.000 description 1
- 208000017015 squamous papilloma Diseases 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
- 238000012800 visualization Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01L—SEMICONDUCTOR DEVICES NOT COVERED BY CLASS H10
- H01L23/00—Details of semiconductor or other solid state devices
- H01L23/28—Encapsulations, e.g. encapsulating layers, coatings, e.g. for protection
- H01L23/31—Encapsulations, e.g. encapsulating layers, coatings, e.g. for protection characterised by the arrangement or shape
- H01L23/3107—Encapsulations, e.g. encapsulating layers, coatings, e.g. for protection characterised by the arrangement or shape the device being completely enclosed
- H01L23/315—Encapsulations, e.g. encapsulating layers, coatings, e.g. for protection characterised by the arrangement or shape the device being completely enclosed the encapsulation having a cavity
-
- 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
-
- 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
- G01N1/38—Diluting, dispersing or mixing 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
- G01N1/36—Embedding or analogous mounting of samples
- G01N2001/362—Embedding or analogous mounting of samples using continuous plastic film to mount sample
Abstract
본 발명은 복수의 격막을 포함하는 다중격막 커버씰에 있어서, 상기 격막 사이에 혼성화 반응에서 발생할 수 있는 격막 간의 용액 누수를 방지하기 위한 진공 흡착홈이 설계된 것을 특징으로 하는 다중격막 커버씰에 관한 것이다.The present invention relates to a multi-diaphragm cover seal comprising a plurality of diaphragm cover seals, wherein a vacuum adsorption groove is designed to prevent leakage of a solution between diaphragms that may occur in a hybridization reaction between the diaphragms. .
본 발명에 따르면, 핵산(DNA 또는 RNA), 핵산유도체, 단백질, 세포, 조직 등의 혼성화 반응 등에 사용되는 커버씰(coverseal)에 진공흡착홈을 설계함으로써 유리슬라이드와의 흡착력을 높이고 격막 간의 용액 누수현상을 방지하며, 설령 격막에서 스며나올 수도 있는 용액은 격막 사이의 진공흡착홈에 흡수되게 함으로써 격막 간의 용액 누수를 완전히 차단할 수 있다. 또한, 본 발명의 다중격막 커버씰은 단일 유리슬라이드 상에서 핵산, 핵산유도체, 단백질, 세포, 조직 등의 혼성화 반응을 위한 마이크로어레이(microarray)용으로도 사용가능하다.According to the present invention, a vacuum adsorption groove is designed in a cover seal used for hybridization of nucleic acids (DNA or RNA), nucleic acid derivatives, proteins, cells, tissues, etc., thereby increasing the adsorption force with the glass slide and leaking solution between the diaphragms. It is possible to prevent the phenomenon, even if the solution that may leak out of the diaphragm to be absorbed in the vacuum suction groove between the diaphragm can completely block the leakage of the solution between the diaphragm. In addition, the multi-diaphragm cover seal of the present invention can also be used for microarrays for hybridization of nucleic acids, nucleic acid derivatives, proteins, cells, tissues, etc. on a single glass slide.
Description
본 발명은 복수의 격막을 포함하는 다중격막 커버씰에 관한 것으로, 더욱 구체적으로는 상기 격막 사이에 혼성화 반응에서 발생할 수 있는 격막 간의 용액 누수를 방지하기 위한 진공 흡착홈이 설계된 것을 특징으로 하는 다중격막 커버씰에 관한 것이다.The present invention relates to a multi-diaphragm cover seal including a plurality of diaphragms, and more specifically, a multi-diaphragm, characterized in that a vacuum adsorption groove is designed to prevent solution leakage between diaphragms that may occur in a hybridization reaction between the diaphragms. It relates to a cover seal.
일반적으로, 커버씰은 유리슬라이드 위에서 핵산(DNA 또는 RNA), 핵산유도체의 염기쌍 형성, 단백질의 항원-항체 반응, 세포(Cell) 또는 조직(tissue)의 형광염색과 현미경을 통한 이미지 분석 등의 각종 혼성화 반응과 형광염색 실험에서 커버글라스 역할로 사용되고 있으며, 이러한 커버씰을 사용함으로써 얻을 수 있는 장점은 유리슬라이드 위에 스포팅(spotting)된 두 종이상의 핵산 또는 단백질 등을 혼성화 반응에 필요한 일정 온도로 유지시킬 수 있으며, 완충용액(buffer)의 증발을 방지하여 적은 양으로도 효과적인 반응을 유도할 수 있다는 것이다(Pasotre, Joseph, N., C. Clampett, J. Miller, K. Porter and D. Miller, A Slide-Incubation Chamber Improves Microwave Assisted Immunostaining. The Journal of Histotechnology 18:115-117, 1995).In general, the cover seal is a variety of nucleic acids (DNA or RNA), base pair formation of nucleic acid derivatives, antigen-antibody reaction of proteins, fluorescence staining of cells or tissues and image analysis through a microscope It is used as a cover glass in the hybridization and fluorescence staining experiments. The advantage of using such cover seals is that two kinds of spotted nucleic acids or proteins on glass slides can be maintained at a certain temperature necessary for the hybridization reaction. It is possible to induce an effective reaction in a small amount by preventing evaporation of the buffer (Pasotre, Joseph, N., C. Clampett, J. Miller, K. Porter and D. Miller, A). Slide-Incubation Chamber Improves Microwave Assisted Immunostaining.The Journal of Histotechnology 18: 115-117, 1995).
최근에는, 각 방(cell)마다 여러 다른 종의 혼성화 반응을 단일 유리슬라이드 위에서 동시에 수행하기 할 수 있도록 복수의 격막을 포함하는 다중격막 커버씰이 사용되고 있다 (McGrath, Charles,M., J.L. Grudzien and A.J. Levine, High-Definition Cell Analysis In Situ Using Microporous Films. CellVision 2:499-509, 1995).Recently, multiple diaphragm cover seals have been used (McGrath, Charles, M., JL Grudzien and multiple diaphragm cover seals) that allow multiple cells to simultaneously perform hybridization reactions of different species on a single glass slide. AJ Levine, High-Definition Cell Analysis In Situ Using Microporous Films.CellVision 2: 499-509, 1995).
그러나, 종래에 사용되고 있는 커버씰은 위에서 언급한 바와 같이 단지 혼성화 반응시 사용되는 완충용액의 증발방지를 통한 사용량의 최소화의 의미 이상으로는 발전되지 못했다. 또한, 다중격막 커버씰의 경우 실험중 격막 간 완충용액의 누수가 가능하므로 방(cell)안의 반응조건이 오염되어 정확한 반응 데이터를 얻을 수 없는 문제점이 있기 때문에, 동시에 다중 반응관찰을 원하는 사용자들에게는 선호되고 있지 않은 실정이다.However, the cover seals used in the related art have not been developed beyond the meaning of minimizing the amount of use by preventing evaporation of the buffer solution used in the hybridization reaction as mentioned above. In addition, in the case of the multi-diaphragm cover seal, the leakage of the buffer solution between the diaphragm is possible during the experiment, so that the reaction conditions in the cell are contaminated, so that accurate reaction data cannot be obtained. It is not preferred.
이에, 본 발명자들은 다중격막 커버씰의 격막 사이에 진공 흡착홈을 설계함으로써 유리슬라이드와의 흡착력을 높일 뿐만 아니라, 혼성화 반응 등에서 발생할 수 있는 격막 간의 용액 누수를 효과적으로 방지하기 할 수 있음을 발견하여 본 발명을 완성하기에 이르렀다.Accordingly, the present inventors have found that by designing a vacuum adsorption groove between the diaphragms of the multi-diaphragm cover seal, it is possible not only to increase the adsorption force with the glass slide, but also to effectively prevent the leakage of solution between the diaphragms that may occur in a hybridization reaction. The invention has been completed.
따라서, 본 발명의 목적은 종래의 다중격막 커버씰의 문제점인 각종 혼성화 반응시 격막간의 용액 누수를 효과적으로 배제할 수 있고, 또한 핵산, 핵산유도체, 단백질, 세포 또는 조직의 혼성화 반응과 형광염색뿐 만 아니라 위 실험의 마이크로어레이(microarray)용으로도 사용가능하게 고려하여 동시에 다중 반응관찰이 용이한 진보된 커버씰을 제공하기 위한 것이다.Accordingly, an object of the present invention can effectively exclude the leakage of solution between the diaphragm during various hybridization reactions, which is a problem of the conventional multi-diaphragm cover seal, and also only hybridization reaction and fluorescence staining of nucleic acid, nucleic acid derivative, protein, cell or tissue. In addition, the present invention is intended to provide an advanced cover seal that can be easily used for the microarray of the above experiments, so that multiple reaction observations can be performed at the same time.
도 1은 실리콘 틀을 도시한 정면도이고,1 is a front view showing a silicon mold,
도 2는 실리콘 틀을 도시한 측면도이고,2 is a side view showing a silicon mold,
도 3은 진공 흡착홈을 확대 도시한 측면도이고,3 is an enlarged side view of a vacuum suction groove;
도 4는 투명 필름판을 도시한 정면도이고,4 is a front view showing a transparent film plate,
도 5는 투명 필름판을 도시한 측면도이고,5 is a side view showing a transparent film plate,
도 6은 다중격막 진공차폐 커버씰 전체 모형도이다.Figure 6 is a multi-diaphragm vacuum shield cover seal overall model.
< 도면의 주요부분에 대한 부호의 설명 ><Description of Symbols for Major Parts of Drawings>
1,2,3,4. 커버씰(cover seal)의 각 방(cell)1,2,3,4. Each cell of the cover seal
5. 진공 흡착홈 6. 실리콘 틀5. Vacuum adsorption groove 6. Silicone mold
7,8,9,10 각 방(cell)의 창(window)7,8,9,10 windows of each cell
11. 완충용액(buffer) 주입구 12. 실리콘 틀과의 접착부분11. Buffer inlet 12. Adhesive part with silicone mold
13. 손잡이13. Handle
상기 목적을 달성하기 위하여, 본 발명은 복수의 격막으로 이루어진 다중격막 커버씰에 있어서, 상기 격막 사이에 진공 흡착홈이 설계된 것을 특징으로 하는 다중격막 커버씰을 제공한다.In order to achieve the above object, the present invention provides a multi-diaphragm cover seal, characterized in that the vacuum adsorption groove is designed between the multi-diaphragm cover seal consisting of a plurality of diaphragms.
바람직하게는, 본 발명은 상기 커버씰이 진공 흡착홈이 설계된 실리콘 틀과이에 부착된 투명필름으로 구성되는 것을 특징으로 하는 다중격막 커버씰을 제공한다. 본 발명에서, 실리콘 틀은 유리슬라이드와 특수한 점착성을 가지고 있으며 혼성화 과정전후 탈부착이 가능하며 온도저항성이 있다. 또한, 투명필름은 방(cell)내의 반응을 관찰할 수 있을 정도의 투명도를 가진다.Preferably, the present invention provides a multi-diaphragm cover seal, characterized in that the cover seal is composed of a transparent film attached to the silicon mold and the vacuum suction groove is designed. In the present invention, the silicone mold has a special adhesiveness with the glass slide and is detachable before and after the hybridization process and has temperature resistance. In addition, the transparent film has a transparency enough to observe the reaction in the cell (cell).
구체적으로, 본 발명에서는 실리콘의 물성을 이용하여 격막 가운데 진공 흡착홈을 냄으로써 커버씰에 압력을 가할 시 유리슬라이드와의 흡착력을 높일 뿐만 아니라, 진공 흡착홈내의 공기가 빠져나와 내부를 진공상태로 만든다. 이로 인해, 혼성화 반응중 발생하는 다중 격막간의 용액 누수, 즉 유리슬라이드와 실리콘 격막 사이의 미세 공간으로 스며나오는 용액이 모두 격막 사이에 가로질러진 진공 흡착홈에 빨려들게 된다.Specifically, in the present invention, by applying the vacuum adsorption groove in the diaphragm by using the physical properties of the silicone to increase the adsorption power with the glass slide when pressure is applied to the cover seal, the air in the vacuum adsorption groove escapes to make the interior into a vacuum state. . As a result, the solution leakage between the multiple diaphragms generated during the hybridization reaction, that is, all the solution leaking into the microcavity between the glass slide and the silicon diaphragm, is sucked into the vacuum adsorption groove intersected between the diaphragms.
본 발명에서, 진공 흡착홈의 길이는 방(cell)의 세로길이보다 긴 것이 바람직하며, 따라서 누수된 어떤 용액도 다른 방(cells)으로의 전이가 발생치 않으므로 누수에 의한 오염 문제는 고려치 않아도 된다.In the present invention, the length of the vacuum adsorption groove is preferably longer than the longitudinal length of the cell, and therefore, any leakage of the solution does not occur in other cells, so the contamination problem due to leakage does not need to be considered. do.
본 발명에서, 투명필름의 각 방에는 상단과 하단에 각각 완충용액 주입구가 형성되어 있는 것이 바람직하며, 이 주입구를 통해 각 방의 반응조건에 맞는 완충용액을 주입할 수 있다.In the present invention, each room of the transparent film is preferably formed with a buffer solution inlet at the top and the bottom, respectively, through the injection port can be injected buffer solution for the reaction conditions of each room.
본 발명에서, 투명필름의 한쪽 단부에는 손잡이가 달린 것이 바람직하며, 이는 혼성화 과정전후에 커버씰의 탈부착을 용이하게 할 수 있다.In the present invention, one end of the transparent film is preferably equipped with a handle, which can facilitate the detachment of the cover seal before and after the hybridization process.
본 발명의 다중격막 커버씰은 각 방(cell)마다 서로 다른 종의 반응을 필요로 하는 어떤 다중반응의 커버씰에도 사용될 수 있으며, 예를 들면, 핵산, 핵산유도체, 단백질, 세포 또는 조직의 혼성화 반응에 사용될 수도 있다(McGrath, Charles, M., J.L. Grudzien and A.J. Levine, Influence of Surface:Volume Ratio of Reaction Chambers on Stoichiometry of Antibody-Based Reactions In Situ. CellVision 2:165-169. 1995., Mantyjarvi M, Syrjanen S, Kaipiainen S, Mantyjarvi R, Kahols T, Syrjanen K, Detection of human papillomavitus type 11 DNA in a conjunctival squamous cell papilloma by in situ hybridization with biothinylated probes. Acta Ophthalmol(Copenh) 67(4):425-429. 1989.).The multi-diaphragm cover seal of the present invention can be used for any multi-reaction cover seal that requires a different species of reaction in each cell, for example, hybridization of nucleic acids, nucleic acid derivatives, proteins, cells or tissues. It can also be used for reactions (McGrath, Charles, M., JL Grudzien and AJ Levine, Influence of Surface: Volume Ratio of Reaction Chambers on Stoichiometry of Antibody-Based Reactions In Situ. Cell Vision 2: 165-169. 1995., Mantyjarvi M , Syrjanen S, Kaipiainen S, Mantyjarvi R, Kahols T, Syrjanen K, Detection of human papillomavitus type 11 DNA in a conjunctival squamous cell papilloma by in situ hybridization with biothinylated probes.Acta Ophthalmol (Copenh) 67 (4): 425-429 1989.).
또한, 본 발명은 핵산, 핵산유도체, 단백질, 세포 또는 조직의 마이크로어레이(microarray)용으로 사용될 수도 있다(David j. Duggan, Michael Bittner, Yidong Chen, Paul Meltzer Jeffery M. Trent. Nature genetics 21(no.1):10-14. 1999., Nicole L.W. van Hal, Oscar Vorst, Adele M.M.L. van Houwelingen, Esther J.Kok, Ad Peijnenburg, Asaph Aharoni, Arjen J. van Tunen, Jaap Keijer. The application of DNA microarrays in gene expression analysis. Journal of Biotechnology 78:271-280. 2000.).The invention may also be used for microarrays of nucleic acids, nucleic acid derivatives, proteins, cells or tissues (David j. Duggan, Michael Bittner, Yidong Chen, Paul Meltzer Jeffery M. Trent. Nature genetics 21 (no .1): 10-14.1999., Nicole LW van Hal, Oscar Vorst, Adele MML van Houwelingen, Esther J.Kok, Ad Peijnenburg, Asaph Aharoni, Arjen J. van Tunen, Jaap Keijer.The application of DNA microarrays in gene expression analysis.Journal of Biotechnology 78: 271-280. 2000.).
또한, 본 발명은 핵산, 핵산유도체, 단백질, 세포 또는 조직의 형광염색에 사용될 수도 있다(Pasotre, Joseph, N., C. Clampett, J. Miller, K. Porter and D. Miller, A Slide-Incubation Chamber Improves Microwave Assisted Immunostaining The Journal of Histotechnology 18:115-117, 1995., Baker-Cairns, B., K. Meyers, R. Hamilton, C. Smith and C. Tornatore Immunohistochemical Staining of Fixed Tissue Using Antigen Retrieval and aThermal Cycler. BioTechniques 20:641-650, 1996., Tarasova N.I., R.. Stauer, J.K. Choi, E.A. Hudson, G. Czerwinski, J.L. Miller, G.N. Pavlakis, C.J. Michejda and S.A. Wank, Visualization of G protein-coupled receptor trafficking with the aid of the green fluorescent protein. Endocytosis and recycling of cholecystokinin receptor type A. J. Biol. Chem. Jun 6;272(23):14817-14824, 1997.).The invention may also be used for fluorescence staining of nucleic acids, nucleic acid derivatives, proteins, cells or tissues (Pasotre, Joseph, N., C. Clampett, J. Miller, K. Porter and D. Miller, A Slide-Incubation Chamber Improves Microwave Assisted Immunostaining The Journal of Histotechnology 18: 115-117, 1995., Baker-Cairns, B., K. Meyers, R. Hamilton, C. Smith and C. Tornatore Immunohistochemical Staining of Fixed Tissue Using Antigen Retrieval and aThermal Cycler.BioTechniques 20: 641-650, 1996., Tarasova NI, R .. Stauer, JK Choi, EA Hudson, G. Czerwinski, JL Miller, GN Pavlakis, CJ Michejda and SA Wank, Visualization of G protein-coupled receptor trafficking with the aid of the green fluorescent protein.Endocytosis and recycling of cholecystokinin receptor type AJ Biol. Chem. Jun 6; 272 (23): 14817-14824, 1997.).
또한, 본 발명은 기타 핵산, 핵산유도체, 단백질, 세포 또는 조직의 산화방지, 보관 또는 완충용액의 증발방지에도 사용될 수 있다.The present invention can also be used to prevent oxidation, storage or buffering of other nucleic acids, nucleic acid derivatives, proteins, cells or tissues.
이하, 본 발명의 바람직한 실시예를 첨부한 도면을 참조하여 설명하면 다음과 같다. 그러나 이것은 예시에 불과할 뿐이며 본 발명의 범위가 실시예 및 도면에 나타난 커버씰에만 한정된 것은 아니다.Hereinafter, with reference to the accompanying drawings, preferred embodiments of the present invention will be described. However, this is only an example and the scope of the present invention is not limited to the cover seals shown in the embodiments and the drawings.
도 1은 본 발명의 실리콘 틀의 정면도로서 실리콘 판에 4개의 방(1,2,3,4)을 구성하도록 디자인되었으며 투명필름으로 덮혀 지지되고있다. 실리콘 틀(6) 가운데 직사각형 형태의 진공 흡착홈(5)이 외부격막(방과 외부 사이)과 내부격막(방과 방 사이)에 파져있다. 이는 커버씰에 압력을 가할 시 유리슬라이드와의 진공상태를 만들어 접착력을 더 높이게 만들기 위함이다. 또한 격막 간 내부 진공 흡착홈(5)은 내부 격막보다 길이가 길며 이는 용액누수가 발생할 시 용액이 내부 진공 흡착홈(5)으로 빨려들게 하여 각 방(1,2,3,4)마다의 용액에 의한 오염을 방지하기 위한 안정장치이기도 하다.1 is a front view of the silicone mold of the present invention, which is designed to configure four chambers 1, 2, 3, and 4 on a silicon plate and is covered with a transparent film. A rectangular vacuum suction groove 5 in the silicon mold 6 is dug in the outer diaphragm (between the room and the outside) and the inner diaphragm (between the room and the room). This is to make the adhesive force higher by making a vacuum with the glass slide when applying pressure to the cover seal. In addition, the inner vacuum adsorption groove 5 between the diaphragms is longer than the inner diaphragm, which causes the solution to be sucked into the inner vacuum adsorption groove 5 when a solution leak occurs, so that the solution of each room (1, 2, 3, 4) is different. It is also a stabilizer to prevent contamination by water.
도 2는 본 발명의 실리콘 틀로의 측면도로서 직사각형 점선은 가로 진공 흡착홈(5)이며 반원형은 세로 진공 흡착홈(5)의 단면부이다. 실리콘 틀의 내부와 외부 가운데 진공 흡착홈(5)을 판다.Fig. 2 is a side view of the silicon mold of the present invention in which a rectangular dotted line is a horizontal vacuum suction groove 5 and a semicircle is a cross section of the vertical vacuum suction groove 5. Dig a vacuum suction groove (5) between the inside and outside of the silicone mold.
도 3은 본 발명의 진공 흡착홈(5)의 단면부를 확대도시한 측면도이다. 이때 진공 흡착홈은 반원통형이다.3 is an enlarged side view of the cross section of the vacuum suction groove 5 of the present invention. At this time, the vacuum suction groove is semi-cylindrical.
도 4는 본 발명의 투명필름의 정면도로서 바깥 테두리 구역은 접착제가 도포되는 영역(12)을 표시한 것이다. 투명필름에 4개의 방(7,8,9,10)은 크기와 모양이 도 1의 실리콘 틀의 4개방(1,2,3,4)과 일치하게 설계하였으며 각 방(7,8,9,10)에는 완충용액 주입구(11)가 각각 상단과 하단에 원형으로 나있다. 이를 통해 완충용액이 주입된다. 손잡이(13)은 혼합화 과정전후의 커버씰 탈부착을 용이케 한다.Fig. 4 is a front view of the transparent film of the present invention, and the outer border zone shows the area 12 where the adhesive is applied. The four rooms (7,8,9,10) on the transparent film are designed to match the size and shape of the four rooms (1,2,3,4) of the silicone mold of FIG. In the 10, the buffer inlet 11 is formed in a circular shape at the top and bottom, respectively. This injects the buffer solution. The handle 13 facilitates detachment of the cover seal before and after the mixing process.
도 5는 본 발명의 투명필름의 측면도로서 접착제 도포부분을 깊게 홈을 내어 실리콘 틀(6)과 투명필름과의 접착부분(12)에만 접착제가 붙도록 고안하였으며 또한 혼성화 반응시 유리 슬라이드에 심어진 핵산, 핵산유도체, 단백질, 세포, 조직들과의 오염을 제거토록 하였다. 투명필름의 두께는 실리콘 틀 두께보다 작으며 필름의 두께에 따라 방(cell)의 부피가 조정되어 진다.Figure 5 is a side view of the transparent film of the present invention designed to attach the adhesive only to the adhesive portion 12 of the silicone mold 6 and the transparent film deeply grooved adhesive coating portion and also the nucleic acid planted in the glass slide during the hybridization reaction To eliminate contamination with nucleic acid derivatives, proteins, cells, and tissues. The thickness of the transparent film is smaller than the thickness of the silicone mold and the volume of the cell is adjusted according to the thickness of the film.
도 6은 본 발명의 다중격막 커버씰의 최종 모형으로서 도 5의 투명필름 판과 도1의 실리콘 틀을 접착하여 만든다. 이때 먼저 실리콘 틀과 투명필름과의 접착부분(12)에 접착제를 붓으로 얇게 도포후 60℃에서 15분 정도 열풍건조후 접착하여 120℃에서 1.5시간에서 2시간 정도 건조시킨다. 얇은 유리면으로 아래위로 가볍게 눌러주면 효과적으로 접착이 된다.FIG. 6 is a final model of the multi-diaphragm cover seal of the present invention made by bonding the transparent film plate of FIG. 5 and the silicon mold of FIG. At this time, first, the adhesive is applied to the adhesive part 12 of the silicone mold and the transparent film thinly with a brush, and then hot-air-dried at 60 ° C. for 15 minutes and then bonded at 120 ° C. for 1.5 hours to 2 hours. A thin glass surface gently presses up and down for effective adhesion.
따라서, 본 발명 이전에 있어서는 격막 간의 용액누수에 의한 오염도 문제때문에 사용을 기피하고 있는 일반적인 다중격막 커버씰의 보다 효과적인 개선안으로서 격막간 진공 흡착홈(5)을 설계하여 유리슬라이드와의 흡착력을 높이고 용액누수에 의한 오염을 방지할 수 있었다. 또한 본 다중격막 커버씰은 핵산, 핵산유도체, 단백질, 세포, 조직등의 혼성화 반응과 형광염색뿐만 아니라 이들 실험의 마이크로어레이(microarray)용도로도 스포팅(spotting)이 가능하게 디자인하였다.Therefore, prior to the present invention, a vacuum adsorption groove (5) between the diaphragms is designed to increase the adsorption force with the glass slide as a more effective improvement of the general multi-diaphragm cover seal, which is avoided due to the problem of contamination due to the leakage of solution between the diaphragms. It was possible to prevent contamination due to leakage. In addition, the multi-diaphragm cover seal was designed to be spotted for hybridization and fluorescence staining of nucleic acids, nucleic acid derivatives, proteins, cells, and tissues, as well as for microarray use of these experiments.
이러한 다중격막 커버씰은 서로 다른 상태의 유전자 발현을 한 유리슬라이드 상에서 또는 동시에 동일한 시료 여러 개에 다른 반응조건을 가하여 한 유리슬라이드 상에서 실험되도록 하여 각 단계별 반복 처리절차를 축소시켜 관찰, 정확한 결과를 얻는 방법에 적용될 수 있다.These multi-diaphragm cover seals are tested on one glass slide with different expression of genes on one glass slide or by applying different reaction conditions to several same samples at the same time. Applicable to the method.
이상에서와 같이 본 발명에 의하면, 다중격막 커버씰의 격막 사이에 진공 흡착홈을 설계함으로써 격막 간의 용액누수와 이에 따른 오염을 방지할 수 있고, 따라서 다중격막 커버씰의 장점인 유전자 발현의 각 단계별 반복 처리절차를 단일 유리슬라이드 내에 축소시켜 관찰하거나 또한 동일한 여러 개의 시료(sample)에 동시적인 다른 반응조건 실험하는데 효과적으로 사용될 수 있다As described above, according to the present invention, by designing the vacuum adsorption groove between the diaphragm of the multi-diaphragm cover seal, it is possible to prevent the leakage of the solution between the diaphragm and contaminating accordingly, and thus, each step of gene expression which is an advantage of the multi-diaphragm cover seal. Repeated procedures can be used to reduce observations in a single glass slide or to effectively test different reaction conditions simultaneously on the same sample.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| KR100773561B1 (en) | 2006-11-07 | 2007-11-05 | 삼성전자주식회사 | Apparatus and method for reducing non-specific amplification in multiplex pcr |
| KR101056581B1 (en) * | 2009-12-18 | 2011-08-11 | 서울시립대학교 산학협력단 | Automated detection system for stable behavior reaction of linear pretty nematodes and similar species under stable conditions by blocking leakage of volatile toxic compounds |
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| US4114611A (en) * | 1977-05-09 | 1978-09-19 | Lyle Judge M | Traction device |
| US4143785A (en) * | 1978-03-16 | 1979-03-13 | Sun Coast Plastic Closures, Inc. | Plastic vacuum sealing cap |
| JPH0334364A (en) * | 1989-06-06 | 1991-02-14 | Natl Semiconductor Corp <Ns> | Complementary pnp/npn ic power transistor |
| JPH0829593A (en) * | 1994-07-20 | 1996-02-02 | Mitsubishi Materials Corp | Sealing structure and sealing member at joint of hermetically sealed container |
| JPH09281106A (en) * | 1996-04-10 | 1997-10-31 | Dainippon Ink & Chem Inc | Plate for inspection and inspecting method |
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| US4114611A (en) * | 1977-05-09 | 1978-09-19 | Lyle Judge M | Traction device |
| US4143785A (en) * | 1978-03-16 | 1979-03-13 | Sun Coast Plastic Closures, Inc. | Plastic vacuum sealing cap |
| JPH0334364A (en) * | 1989-06-06 | 1991-02-14 | Natl Semiconductor Corp <Ns> | Complementary pnp/npn ic power transistor |
| JPH0829593A (en) * | 1994-07-20 | 1996-02-02 | Mitsubishi Materials Corp | Sealing structure and sealing member at joint of hermetically sealed container |
| JPH09281106A (en) * | 1996-04-10 | 1997-10-31 | Dainippon Ink & Chem Inc | Plate for inspection and inspecting method |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100773561B1 (en) | 2006-11-07 | 2007-11-05 | 삼성전자주식회사 | Apparatus and method for reducing non-specific amplification in multiplex pcr |
| KR101056581B1 (en) * | 2009-12-18 | 2011-08-11 | 서울시립대학교 산학협력단 | Automated detection system for stable behavior reaction of linear pretty nematodes and similar species under stable conditions by blocking leakage of volatile toxic compounds |
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