JP4374435B2 - Tissue embedding method with highly hydrophilic polymer. - Google Patents
Tissue embedding method with highly hydrophilic polymer. Download PDFInfo
- Publication number
- JP4374435B2 JP4374435B2 JP2004220045A JP2004220045A JP4374435B2 JP 4374435 B2 JP4374435 B2 JP 4374435B2 JP 2004220045 A JP2004220045 A JP 2004220045A JP 2004220045 A JP2004220045 A JP 2004220045A JP 4374435 B2 JP4374435 B2 JP 4374435B2
- Authority
- JP
- Japan
- Prior art keywords
- tissue
- highly hydrophilic
- hydrophilic monomer
- hydrophilic polymer
- polymerization
- 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.)
- Expired - Lifetime
Links
- 238000000034 method Methods 0.000 title claims description 18
- 229920001477 hydrophilic polymer Polymers 0.000 title claims description 11
- 238000006116 polymerization reaction Methods 0.000 claims description 23
- 239000000178 monomer Substances 0.000 claims description 21
- 239000000243 solution Substances 0.000 claims description 13
- 239000003795 chemical substances by application Substances 0.000 claims description 8
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 6
- 239000003431 cross linking reagent Substances 0.000 claims description 6
- 229910001873 dinitrogen Inorganic materials 0.000 claims description 6
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 4
- 239000002504 physiological saline solution Substances 0.000 claims description 3
- 238000010526 radical polymerization reaction Methods 0.000 claims description 3
- SWXVUIWOUIDPGS-UHFFFAOYSA-N diacetone alcohol Natural products CC(=O)CC(C)(C)O SWXVUIWOUIDPGS-UHFFFAOYSA-N 0.000 claims description 2
- 235000019441 ethanol Nutrition 0.000 claims description 2
- 238000007654 immersion Methods 0.000 claims 1
- 239000003505 polymerization initiator Substances 0.000 claims 1
- 210000001519 tissue Anatomy 0.000 description 37
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 12
- 150000003254 radicals Chemical class 0.000 description 10
- 230000000977 initiatory effect Effects 0.000 description 9
- MYRTYDVEIRVNKP-UHFFFAOYSA-N 1,2-Divinylbenzene Chemical compound C=CC1=CC=CC=C1C=C MYRTYDVEIRVNKP-UHFFFAOYSA-N 0.000 description 7
- 210000004027 cell Anatomy 0.000 description 7
- QIGBRXMKCJKVMJ-UHFFFAOYSA-N Hydroquinone Chemical compound OC1=CC=C(O)C=C1 QIGBRXMKCJKVMJ-UHFFFAOYSA-N 0.000 description 4
- 229920000642 polymer Polymers 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 230000005587 bubbling Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 230000007613 environmental effect Effects 0.000 description 3
- 230000003834 intracellular effect Effects 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- FSWDLYNGJBGFJH-UHFFFAOYSA-N n,n'-di-2-butyl-1,4-phenylenediamine Chemical compound CCC(C)NC1=CC=C(NC(C)CC)C=C1 FSWDLYNGJBGFJH-UHFFFAOYSA-N 0.000 description 3
- 239000002861 polymer material Substances 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 239000011347 resin Substances 0.000 description 3
- 229920005989 resin Polymers 0.000 description 3
- 238000006467 substitution reaction Methods 0.000 description 3
- 239000002344 surface layer Substances 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 2
- 230000008595 infiltration Effects 0.000 description 2
- 238000001764 infiltration Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- ZIUHHBKFKCYYJD-UHFFFAOYSA-N n,n'-methylenebisacrylamide Chemical compound C=CC(=O)NCNC(=O)C=C ZIUHHBKFKCYYJD-UHFFFAOYSA-N 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 230000008520 organization Effects 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 230000008961 swelling Effects 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 239000002699 waste material Substances 0.000 description 2
- VUZNLSBZRVZGIK-UHFFFAOYSA-N 2,2,6,6-Tetramethyl-1-piperidinol Chemical compound CC1(C)CCCC(C)(C)N1O VUZNLSBZRVZGIK-UHFFFAOYSA-N 0.000 description 1
- SMZOUWXMTYCWNB-UHFFFAOYSA-N 2-(2-methoxy-5-methylphenyl)ethanamine Chemical compound COC1=CC=C(C)C=C1CCN SMZOUWXMTYCWNB-UHFFFAOYSA-N 0.000 description 1
- NIXOWILDQLNWCW-UHFFFAOYSA-N 2-Propenoic acid Natural products OC(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 description 1
- 229940044192 2-hydroxyethyl methacrylate Drugs 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- WOBHKFSMXKNTIM-UHFFFAOYSA-N Hydroxyethyl methacrylate Chemical compound CC(=C)C(=O)OCCO WOBHKFSMXKNTIM-UHFFFAOYSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 230000002421 anti-septic effect Effects 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 230000020169 heat generation Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 210000004738 parenchymal cell Anatomy 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Landscapes
- Sampling And Sample Adjustment (AREA)
- Polymerisation Methods In General (AREA)
Description
本発明は、高親水性高分子による組織包埋方法に関するものである。 The present invention relates to a tissue embedding method using a highly hydrophilic polymer.
電子顕微鏡の試料作製の薄切りや、実習教材のプラスチネーション作製などを目的として、従来からも凍結切片作成法や、各種樹脂への包埋等のプロセスが開発され、日常的に用いられている。
主に耐久性確保の観点から開発された各種樹脂への包埋プロセスでは、組織は元来水分を多く含むことから、樹脂材料として一般に親水性の低い高分子材料を使用している。この親水性の低い高分子材料は、組織との親和性が低いため、該包埋プロセスは、組織からの真空脱水や置換といった工程が組み込まれる。
Processes such as frozen section preparation and embedding in various resins have been developed and used on a daily basis for the purpose of slicing specimens for electron microscopes and plastination for training materials.
In the embedding process in various resins developed mainly from the viewpoint of ensuring durability, since the tissue originally contains a lot of moisture, a polymer material having low hydrophilicity is generally used as the resin material. Since this low hydrophilic polymer material has low affinity with the tissue, the embedding process incorporates steps such as vacuum dehydration and replacement from the tissue.
前記従来の包埋プロセスは、低親水性高分子材料を使用するため、多くの特殊設備と処理工程を必要とし、又取扱者は、高分子に対する専門知識が要求され、処理廃液による環境負荷の問題、人体への毒性の問題、組織への重合時の発熱による影響等を厳重に管理し安全を確保しなければない。また製造コストも嵩む。
そこで本発明者等は、取り扱い上比較的安全な、高親水性高分子の応用に着目した。
しかし、この高親水性高分子のみを組織の包埋プロセスに単に応用したのでは、親水性が高いため組織が膨潤する問題、更に重合速度が早く、大きさにもよるが、組織内全域に浸透する前に、表層部などの初期浸透部のみ重合してしまう等の問題があり実用化できない。
Since the conventional embedding process uses a low hydrophilic polymer material, it requires a lot of special equipment and processing steps, and the operator is required to have expertise in polymers, and the environmental load caused by the processing waste liquid is reduced. We must strictly manage issues, toxicity to humans, and the effects of heat generation during polymerization on tissues to ensure safety. In addition, the manufacturing cost increases.
Therefore, the present inventors paid attention to the application of a highly hydrophilic polymer that is relatively safe in handling.
However, if only this highly hydrophilic polymer is simply applied to the tissue embedding process, the tissue is swollen due to its high hydrophilicity, and the polymerization rate is high, depending on the size. Prior to infiltration, there is a problem that only the initial infiltration part such as the surface layer part is polymerized.
本発明はこれらの問題を解決するためになされたものであり、その特徴とする技術条件は、次の（1）にある。
(1)、細胞内に水分とラジカルソースを含む組織をラジカル重合反応させ包埋標本化するに際して、前記組織を生理食塩水に浸漬して洗浄した後に、高親水性モノマーに強度付与用の架橋剤2〜4w％と重合開始制御剤100〜400ppm含有させ30〜40℃にした高親水性モノマー溶液に浸漬し、この浸漬中に窒素ガスを所定時間バブリングし、その後に静置して組織細胞内のラジカルソースとの重合反応を開始させ組織全体を重合化させ、この後アセトン・エチルアルコールにより洗浄することを特徴とする高親水性高分子による組織包埋方法。
The present invention has been made in order to solve these problems, and the technical conditions characterized by it are the following (1).
(1) When a tissue containing moisture and a radical source in a cell is subjected to a radical polymerization reaction to prepare an embedded sample, the tissue is immersed in physiological saline and washed, and then a highly hydrophilic monomer is crosslinked for strength. Soaked in a highly hydrophilic monomer solution containing 2 to 4 w% of the agent and 100 to 400 ppm of the polymerization initiation control agent and adjusted to 30 to 40 ° C., nitrogen gas was bubbled for a predetermined time during this soaking, and the tissue cells were then left standing A tissue embedding method using a highly hydrophilic polymer, characterized by initiating a polymerization reaction with a radical source therein to polymerize the entire tissue and then washing with acetone / ethyl alcohol .
本発明は、上記簡易な構成により組織を膨潤させることなく組織全域の細胞内水分を高親水性モノマー溶液に置換させ、この後に組織細胞内のラジカルソースによって重合反応を開始させるものであり、これにより、組織全体の重合化を確立したものである。
この方法では、包埋組織についてホルマリン固定等の前処理が必須でないことから、幅広い状態にある組織への応用が可能である。
これにより本発明は、更に次の優れた効果を有する。
(1)、真空ラインなどの実験設備を持たない、医学部等の研究室でも重合が可能である。
(2)、高分子化学の専門知識がなくても、安全に利用できるプロセスである。
(3)、組織への重合による熱の影響のない、低温で重合するプロセスである。
(4)、人体への毒性がほとんど無いため、教材として安全に使用することができる。
(5)、従来プロセスで必要であった、置換溶媒などの処理廃液の排出を激減させ、環境負荷を最小限にすることができる。
The present invention replaces intracellular moisture in the entire tissue with a highly hydrophilic monomer solution without swelling the tissue with the above simple structure, and then initiates a polymerization reaction with a radical source in the tissue cell. Thus, polymerization of the entire tissue is established.
In this method, since pretreatment such as formalin fixation is not essential for the embedded tissue, it can be applied to a wide variety of tissues.
Thereby, this invention has the following outstanding effect further.
(1) Polymerization is possible even in laboratories such as medical schools that do not have experimental facilities such as vacuum lines.
(2) It is a process that can be used safely without specialized knowledge of polymer chemistry.
(3) A process that polymerizes at a low temperature without the influence of heat due to polymerization on the tissue.
(4) Since there is almost no toxicity to the human body, it can be used safely as a teaching material.
(5) It is possible to drastically reduce the discharge of processing waste liquid such as substitution solvent, which was necessary in the conventional process, and to minimize the environmental burden.
本発明の最良の形態例は、細胞内に水分とラジカルソースを含む組織を、該組織からの吸水量と目標とする重合体の硬度等を基準にした割合で架橋剤と重合開始制御剤を配合した前記の高親水性モノマー溶液に浸漬し、窒素ガスバブリング後静置して重合させ、重合後洗浄する高親水性高分子による組織包埋方法である。
高親水性モノマー溶液は、例えば、生理食塩水で洗浄したブタ肝臓5 mm厚の切片の場合、高親水性高分子としてN-vinyl-2-pyrolidoneをベースにし、架橋剤としてN,N'-Methlenebisacrylamideを2〜4 w％、重合開始制御剤としてN,N'-Di-sec-butyl-p-phenylenediamineを100〜400 ppm用いることにより、組織全域の細胞内水分を高親水性モノマー溶液に置換させた後に組織細胞内のラジカルソースとの重合反応を開始させ組織全体の重合化を実現し、上記目標の包埋組織を得た。
In the best mode of the present invention, a tissue containing moisture and a radical source in a cell is mixed with a crosslinking agent and a polymerization initiation control agent at a ratio based on the water absorption from the tissue and the hardness of the target polymer. It is a tissue embedding method using a highly hydrophilic polymer that is immersed in the blended highly hydrophilic monomer solution , allowed to stand after nitrogen gas bubbling, polymerized, and washed after polymerization.
For example, in the case of a slice of pig liver 5 mm thick washed with physiological saline, the highly hydrophilic monomer solution is based on N-vinyl-2-pyrolidone as a highly hydrophilic polymer and N, N'- Replacing intracellular water in the entire tissue with highly hydrophilic monomer solution by using 2-4 w% of Methlenebisacrylamide and 100-400 ppm of N, N'-Di-sec-butyl-p-phenylenediamine as a polymerization initiation controller Then, a polymerization reaction with the radical source in the tissue cells was started to realize polymerization of the entire tissue, and the target embedded tissue was obtained.
而して、本発明において、対象とする細胞内に水分とラジカルソースを含む組織としては、各種動物の各種管空性臓器、筋組織を含む実質性臓器等である。
本発明においては、ホルマリン等による前処理は必須ではない。従って、従来から行われてきた、ホルマリン処理を含むあらゆる組織固定法および未処理組織に適用可能である。しかし、以下のような場合、組織を予めホルマリン固定を行う。
感染性や変質が見られる試料の作成において、ホルマリン処理が適切であると判断される場合。
長期保存を前提として、高い防腐性を持たせたい場合。
また既にホルマリン固定がなされている、過去の保存試料に対しても、本発明を適用することが可能である。
And Thus, in the present invention, as an organization containing moisture and a radical source in the cell of interest, various tubes empty organ of various animals, a parenchymal organ, such as including muscle tissue.
In the present invention, pretreatment with formalin or the like is not essential. Therefore, it can be applied to all tissue fixing methods including formalin treatment and untreated tissue, which have been conventionally performed. However, formalin fixation is performed in advance in the following cases.
When it is judged that formalin treatment is appropriate for the preparation of infectious or altered samples.
When you want to have high antiseptic properties on the premise of long-term storage.
In addition, the present invention can be applied to past stored samples that have already been fixed with formalin.
本発明において、高親水性モノマー（高親水性高分子）とは、N-vinyl-2-pyrolidone、Acrylic acid、2-Hydroxyethylmethacrylate、Vinylalchol polymerの一種を言う、その選定理由は、次の通りである。
入手し易く、比較的安価であること。
化学の専門知識を持たない者でも、安全に扱えること。
著明な毒性がないこと。
環境負荷が小さいこと。
In the present invention, the highly hydrophilic monomer (highly hydrophilic polymer) refers to one of N-vinyl-2-pyrolidone, Acrylic acid, 2-Hydroxyethylmethacrylate, and Vinylalchol polymer. The reason for selection is as follows. .
Easy to get and relatively inexpensive.
Even those without chemical expertise should be able to handle it safely.
There is no significant toxicity.
Environmental impact is small.
本発明において、高親水性モノマー溶液における架橋剤の含有理由と具体例及びその好ましい含有割合範囲は、次の通りである。
理由：高親水性モノマーのみでは、プラスチネーションや切片作製に必要とされる強度が得られない。また、保存時の吸湿による劣化・膨潤や、寸法精度を保つ必要がある。これらを解決する目的で、架橋剤による網目構造の構築が望ましい。
具体例：架橋剤としては、N,N'-Methylenebisacrylamide、Divinylbenzeneなどがある。
架橋剤の具体例個々の含有割合範囲：
1.N,N'-Methylenebisacrylamide:高親水性モノマーに対して0.5〜4.5 wt.%が好ましい。
2.Divinylbenzene: 高親水性モノマーに対して1.0〜6.5 wt.%が好ましい。
In the present invention, the reason for containing the crosslinking agent in the highly hydrophilic monomer solution , specific examples thereof, and preferred content ratio ranges thereof are as follows.
Reason: The strength required for plastination and slice preparation cannot be obtained with only a highly hydrophilic monomer . Moreover, it is necessary to maintain deterioration and swelling due to moisture absorption during storage and dimensional accuracy. In order to solve these problems, it is desirable to construct a network structure using a crosslinking agent.
Specific examples: Examples of the cross-linking agent include N, N'-Methylenebisacrylamide, and divinylbenzene.
Specific content range of specific examples of crosslinking agents:
1. N, N'-Methylenebisacrylamide: 0.5 to 4.5 wt.% Is preferable with respect to the highly hydrophilic monomer .
2.Divinylbenzene: 1.0 to 6.5 wt.% Is preferable with respect to the highly hydrophilic monomer .
本発明において、高親水性モノマー溶液における重合開始制御剤の含有理由と具体例及びその好ましい含有割合範囲は、次の通りである。
理由：組織内にはそのいたるところにラジカルソースが存在する。そのため高親水性モノマーが水と置換された部分から速やかに重合が開始し、表層で重合が進行することで、内部の水分が高親水性モノマーに置換されることの障害となる。重合開始制御剤は、組織全体を均等に全量重合化反応させるために、組織全域の細胞内水分が高親水性モノマー溶液に置換されるまでの間、組織表層部等の初期置換部細胞内のラジカルソースによる重合化反応を抑制するために含有させるものである。
具体例：重合開始制御剤としては、N,N'-Di-sec-butyl-p-phenylenediamine、hydroquinone、2,2,6,6-tetramethylpiperidinooxy, freeradical等がある。濃度は高親水性モノマーに対して100〜400 ppmとすることが好ましい。
重合開始制御剤の具体例個々の含有割合（濃度）については、対象とする組織の種類により、水分量やラジカルソース含有率と置換速度が異なり、また同種でも大きさにより完全置換時間が変化するため、これらの要素と重合開始制御剤の種類に応じて実験的、経験的にその最適濃度範囲を設定することができる。組織のラジカル種の同定や定量は、例えばESRによるspin-trap法を用いるなど、臓器・組織別の最適値を求めることで、より広範な用途に応用することができる。
In the present invention, the reason why the polymerization initiation controller is contained in the highly hydrophilic monomer solution , specific examples thereof, and preferred content ratio ranges thereof are as follows.
Reason: There are radical sources throughout the organization. For this reason, the polymerization starts promptly from the portion where the highly hydrophilic monomer is replaced with water, and the polymerization proceeds on the surface layer, which is an obstacle to the replacement of the internal moisture with the highly hydrophilic monomer . In order to cause the entire tissue to uniformly polymerize the entire tissue, the polymerization initiation control agent is used in the initial replacement part cells such as the tissue surface layer part until the intracellular moisture in the whole tissue is replaced with the highly hydrophilic monomer solution . It is contained in order to suppress a polymerization reaction caused by a radical source.
Specific examples: Examples of the polymerization initiation control agent include N, N'-Di-sec-butyl-p-phenylenediamine, hydroquinone, 2,2,6,6-tetramethylpiperidinooxy, freeradical and the like. The concentration is preferably 100 to 400 ppm with respect to the highly hydrophilic monomer .
Specific examples of the polymerization initiation control agent The content (concentration) of each component varies depending on the type of target tissue, the amount of water, the content of radical source, and the substitution rate. Even in the same type, the complete substitution time varies depending on the size. Therefore, the optimum concentration range can be set experimentally and empirically according to the types of these elements and the polymerization initiation control agent. The identification and quantification of tissue radical species can be applied to a wider range of applications by determining the optimum value for each organ or tissue, for example, using the spin-trap method by ESR.
本発明において、前記高親水性モノマー溶液に組織を浸漬して窒素ガスバブリングする理由と好ましい温度範囲や所要時間範囲は、次の通りである。
理由：本発明の基本反応はラジカル重合反応であることから、酸素除去を目的とする。
方法：室温において、窒素ガスを注射針から高親水性モノマー溶液中に導入する。導入時間は浸漬する溶液の量に応じて0.2 l/min.の流量で5〜15分とする。
本発明において、窒素ガスバブリング後静置する理由と好ましい温度や所要時間は、次の通りである。
理由：高親水性モノマー溶液の組織内への浸透、および重合反応を行うため。
In the present invention, the reason for immersing the tissue in the highly hydrophilic monomer solution and bubbling nitrogen gas and the preferred temperature range and required time range are as follows.
Reason: Since the basic reaction of the present invention is a radical polymerization reaction, the purpose is to remove oxygen.
Method: At room temperature, nitrogen gas is introduced into the highly hydrophilic monomer solution from the injection needle. The introduction time is 5 to 15 minutes at a flow rate of 0.2 l / min. Depending on the amount of the solution to be immersed.
In the present invention, the reason for standing after nitrogen gas bubbling and the preferred temperature and required time are as follows.
Reason: To perform penetration of the highly hydrophilic monomer solution into the tissue and polymerization reaction.
本発明の実施例を、表１〜表５に詳細に紹介する。 Examples of the present invention are introduced in detail in Tables 1-5.
St:Stylene
MBA:Methlene
DBPD:N,N'-Di-sec-butyl-p-phenylenediamine
HQN:hydroquinone
DVB:Divinylbenzene
St: Stylene
MBA: Methlene
DBPD: N, N'-Di-sec-butyl-p-phenylenediamine
HQN: hydroquinone
DVB: Divinylbenzene
本発明は、前記の効果に詳述のように、人体及び自然環境に優しく、且つ特殊設備を不要とする簡単な処理工程で、誰もが容易に安全に管理し実施することができ、且つ安価な製造方法であり、この種産業の利用可能性は多大なものがある。 As described in detail in the above-mentioned effects, the present invention can be easily and safely managed and implemented by anyone with a simple treatment process that is gentle to the human body and the natural environment and does not require special equipment. It is an inexpensive manufacturing method, and the applicability of this type of industry is enormous.
Claims (1)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2004220045A JP4374435B2 (en) | 2004-07-28 | 2004-07-28 | Tissue embedding method with highly hydrophilic polymer. |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2004220045A JP4374435B2 (en) | 2004-07-28 | 2004-07-28 | Tissue embedding method with highly hydrophilic polymer. |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2006036957A JP2006036957A (en) | 2006-02-09 |
| JP4374435B2 true JP4374435B2 (en) | 2009-12-02 |
Family
ID=35902298
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2004220045A Expired - Lifetime JP4374435B2 (en) | 2004-07-28 | 2004-07-28 | Tissue embedding method with highly hydrophilic polymer. |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP4374435B2 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2015086190A (en) * | 2013-10-31 | 2015-05-07 | 国立大学法人 大分大学 | Infusion solution for cadaver preservation |
Families Citing this family (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP4956839B2 (en) * | 2008-02-13 | 2012-06-20 | 国立大学法人 大分大学 | Tissue embedding method with highly hydrophilic polymer monomer aqueous solution |
| EP3108218A4 (en) * | 2014-02-21 | 2017-11-15 | Massachusetts Institute Of Technology | Expansion microscopy |
| US11408890B2 (en) | 2015-04-14 | 2022-08-09 | Massachusetts Institute Of Technology | Iterative expansion microscopy |
| JP6721899B2 (en) * | 2016-03-30 | 2020-07-15 | 国立大学法人 大分大学 | Embedding method for electron microscope mainly composed of highly hydrophilic polymer monomer |
| WO2018136856A1 (en) | 2017-01-23 | 2018-07-26 | Massachusetts Institute Of Technology | Multiplexed signal amplified fish via splinted ligation amplification and sequencing |
| WO2018157074A1 (en) | 2017-02-24 | 2018-08-30 | Massachusetts Institute Of Technology | Methods for diagnosing neoplastic lesions |
| WO2018157048A1 (en) | 2017-02-24 | 2018-08-30 | Massachusetts Institute Of Technology | Methods for examining podocyte foot processes in human renal samples using conventional optical microscopy |
| US11180804B2 (en) | 2017-07-25 | 2021-11-23 | Massachusetts Institute Of Technology | In situ ATAC sequencing |
| WO2019156957A1 (en) | 2018-02-06 | 2019-08-15 | Massachusetts Institute Of Technology | Swellable and structurally homogenous hydrogels and methods of use thereof |
| WO2020013833A1 (en) | 2018-07-13 | 2020-01-16 | Massachusetts Institute Of Technology | Dimethylacrylamide (dmaa) hydrogel for expansion microscopy (exm) |
| CA3130889A1 (en) | 2019-02-22 | 2020-08-27 | Massachusetts Institute Of Technology | Iterative direct expansion microscopy |
| US12265004B2 (en) | 2019-11-05 | 2025-04-01 | Massachusetts Institute Of Technology | Membrane probes for expansion microscopy |
| US11802822B2 (en) | 2019-12-05 | 2023-10-31 | Massachusetts Institute Of Technology | Multiplexed expansion (MultiExM) pathology |
-
2004
- 2004-07-28 JP JP2004220045A patent/JP4374435B2/en not_active Expired - Lifetime
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2015086190A (en) * | 2013-10-31 | 2015-05-07 | 国立大学法人 大分大学 | Infusion solution for cadaver preservation |
Also Published As
| Publication number | Publication date |
|---|---|
| JP2006036957A (en) | 2006-02-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP4956839B2 (en) | Tissue embedding method with highly hydrophilic polymer monomer aqueous solution | |
| JP4374435B2 (en) | Tissue embedding method with highly hydrophilic polymer. | |
| US6358557B1 (en) | Graft polymerization of substrate surfaces | |
| CN116003692B (en) | Surface graft crosslinked zwitterionic polymer coating, and preparation method and application thereof | |
| EP0068509B1 (en) | Antithrombogenic biomedical material | |
| Turner et al. | Water sorption of poly (methyl methacrylate): 2. Effects of crosslinks | |
| EP3461506B1 (en) | Material for intraocular lenses | |
| WO2019100571A1 (en) | Method for surface hydrophilic modification of silicon rubber and application method | |
| JPH02228309A (en) | Method for modifying surface characteristic of preformed base material by graft polymerization | |
| US10556040B2 (en) | Surface-modified metal and method for modifying metal surface | |
| Omer et al. | Hydrogels from dextran and soybean oil by UV photo‐polymerization | |
| CN111978567A (en) | Poly (2-hydroxyethyl methacrylate)/N-vinyl pyrrolidone binary copolymer hydrogel and preparation method thereof | |
| Razzak et al. | Modification of natural rubber tubes for biomaterials I. Radiation‐induced grafting of N, N‐dimethyl acrylamide onto natural rubber tubes | |
| JPH0311787B2 (en) | ||
| Migliaresi et al. | Water sorption and mechanical properties of 2‐hydroxyethyl‐methacrylate and methylmethacrylate copolymers | |
| JP3314451B2 (en) | Surface treatment method for polymer materials | |
| JP3495802B2 (en) | Medical materials and polyvinyl alcohol polymers having phosphorylcholine groups | |
| El‐Rehim et al. | Properties and biocompatibility of polypropylene graft copolymer films | |
| Iwasaki et al. | Hydrogel‐like elastic membrane consisting of semi‐interpenetrating polymer networks based on a phosphorylcholine polymer and a segmented polyurethane | |
| JPS60242003A (en) | Improved wood | |
| Pinchuk et al. | The interaction of urea with the generic class of poly (2‐hydroxyethyl methacrylate) hydrogels | |
| KR20230160907A (en) | antithrombotic material | |
| Dessouki et al. | Radiation grafting of acrylic and methacrylic acids onto poly (tetrafluoroethylene‐hexafluoropropylene‐vinylidene fluoride) TFB films | |
| JPH11226113A (en) | Blood compatible polyurethane-hydrophilic high polymer blend | |
| Radzali et al. | 60Co-induced grafting of dual polymer (acrylic acid-co-HEMA) onto expanded poly (tetrafluoroethylene) membranes |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20060623 |
|
| A977 | Report on retrieval |
Free format text: JAPANESE INTERMEDIATE CODE: A971007 Effective date: 20080804 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20080812 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20081014 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20081111 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20081125 |
|
| A02 | Decision of refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A02 Effective date: 20090106 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20090204 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20090302 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20090304 |
|
| A911 | Transfer to examiner for re-examination before appeal (zenchi) |
Free format text: JAPANESE INTERMEDIATE CODE: A911 Effective date: 20090401 |
|
| TRDD | Decision of grant or rejection written | ||
| A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20090811 |
|
| A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 |
|
| R150 | Certificate of patent or registration of utility model |
Ref document number: 4374435 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
| EXPY | Cancellation because of completion of term |