JP2019056708A - 膨張顕微鏡法 - Google Patents
膨張顕微鏡法 Download PDFInfo
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Abstract
Description
本願は、その内容が、その全体において参照により本明細書に援用される2014年2月21日に出願された米国仮特許出願番号61/943,045の利益を主張する。
本発明は、国立衛生研究所により認められた助成金番号1DP1NS087724の下、米国政府の支援によりなされた。政府は、本発明に一定の権利を有する。
本発明は、顕微鏡法、特に生物学的標本の光学的画像化に関する。
顕微鏡法は、固定された細胞および組織中の小さな構造の画像を光学的に拡大することにより、価値のある生物学的情報を提供している。しかしながら、かかる画像化技術の解像度は、発光源の波長の約半分に限定される。スペクトルの可視領域において、これは1ミクロンの1/4〜1/3のオーダーである(250〜330nm)。不幸にも、従来の光学的顕微鏡法ではこの長さの尺度未満の非侵襲性の観察が利用できない非常に多くの範囲の生物学的構造/系がある。
本発明は、生物学的標本自体を物理的に膨張することに基づく、古典的な顕微鏡法回析限界よりも良好な解像度での生物学的標本の光学的画像化のための方法である。この方法において、培養された細胞、固定された組織、または原則として、生物学的材料を含む他の型の目的の試料に、組成物または化学カクテルを注入し、該組成物または化学カクテルで試料材料を包埋し、次いで該組成物を、好ましくはナノスケールの正確さで三次元において全方向に等しく膨張させ得る。
〔1〕顕微鏡法のための目的の試料を拡大させるための方法であって、目的の試料を膨潤性(swellable)材料に埋め込む工程、および該材料を膨潤させる工程を含む、方法
に関する。
本発明は、顕微鏡法のための拡大された目的の試料、および目的の試料を拡大するための方法、標本中で膨潤性(swellable)ポリマー網目状組織を合成することによる古典的顕微鏡法回析限界よりも良好な解像度での目的の試料の光学的画像化に関し、標本は、物理的に膨張され、物理的な拡大がもたらされ得る。本発明はまた、目的の試料を拡大するための本明細書に記載される組成物の使用を含む。
図1は、本発明の方法の例示的な態様における工程を示すフローチャートである。
[1]顕微鏡法のための目的の試料を拡大させるための方法であって、目的の試料を膨潤性(swellable)材料に埋め込む工程、および該材料を膨潤させる工程を含む、方法。
[2]該試料が標識される、[1]記載の方法。
[3]該試料が、免疫蛍光染色、免疫組織化学的染色または免疫細胞化学的染色により標識される、[2]記載の方法。
[4]該標識された試料が、膨潤性材料に固定される、[2]記載の方法。
[5]該標識が、膨潤性材料に結合される、[4]記載の方法。
[6]該試料を膨潤性材料に埋め込む工程が、該試料に、膨潤性ポリマーの前駆体を含む組成物を浸透化させること、およびインサイチュで膨潤性ポリマーを形成することを含む、[1]記載の方法。
[7]該組成物が、1つ以上の水溶性モノマー前駆体を含む水溶液である、[6]記載の方法。
[8]該溶液が、アクリレート、アクリルアミドおよびN,N-メチレンビスアクリルアミドを含む、[7]記載の方法。
[9]該膨潤性材料がポリアクリレートである、[1]または[6]記載の方法。
[10]該膨潤性材料が試料中に浸透化される、[1]または[6]記載の方法。
[11]該試料が、膨潤性材料に固定される、[10]記載の方法。
[12]該試料が、水を添加することにより全方向に等しく膨潤され、拡大された試料を生じる、[1]記載の方法。
[13]水の添加により、埋め込まれた試料が元の大きさの4x〜5xに膨張する(expand)、[12]記載の方法。
[14]該試料に1つ以上のモノマー前駆体を浸透化させる前に、該試料を界面活性剤で処理する、[6]記載の方法。
[15]該試料を膨潤させる前に、該試料を酵素消化に供し、内部にある(underlying)内因性生物学的分子を破壊する、[1]記載の方法。
[16]該消化が、標識をそのままで、かつ膨潤性材料に固定させたままにする、[14]記載の方法。
[17]目的の顕微鏡法の拡大可能な試料を調製するための方法であって、目的の試料を膨潤性材料に埋め込むことを含む、方法。
[18]該試料が標識される、[17]記載の方法。
[19]該試料が、免疫蛍光染色、免疫組織化学的染色または免疫細胞化学的染色により標識される、[18]記載の方法。
[20]該標識された試料が、膨潤性材料に固定される、[18]記載の方法。
[21]該標識が、膨潤性材料に結合される、[20]記載の方法。
[22]該試料を膨潤性材料に埋め込む工程が、該試料に、膨潤性ポリマーの前駆体を含む組成物を浸透化させること、およびインサイチュで膨潤性ポリマーを形成することを含む、[17]記載の方法。
[23]該組成物が、1つ以上の水溶性モノマー前駆体を含む水溶液である、[22]記載の方法。
[24]該溶液が、アクリレート、アクリルアミドおよびN,N-メチレンビスアクリルアミドを含む、[23]記載の方法。
[25]膨潤性材料がポリアクリレートである、[17]または[22]記載の方法。
[26]該膨潤性材料が、試料中に浸透化される、[17]または[22]記載の方法。
[27]該試料が、膨潤性材料に固定される、[26]記載の方法。
[28]該試料に1つ以上のモノマー前駆体を浸透化させる前に、該試料を界面活性剤で処理する、[22]記載の方法。
[29]試料の高解像画像を作製するための顕微鏡法の方法であって、目的の試料を拡大する工程および該拡大された試料を顕微鏡下で見分する工程を含む、方法。
[30]目的の試料を拡大する工程が、目的の試料を膨潤性材料に埋め込むことおよび該材料を膨潤させることを含む、[29]記載の方法。
[31]該試料が標識される、[30]記載の方法。
[32]該試料が、免疫蛍光染色、免疫組織化学的染色または免疫細胞化学的染色により標識される、[31]記載の方法。
[33]該標識された試料が、膨潤性材料に固定される、[31]記載の方法。
[34]該標識が、膨潤性材料に結合される、[33]記載の方法。
[35]該試料を膨潤性材料に埋め込む工程が、該試料に、膨潤性ポリマーの前駆体を含む組成物を浸透化させること、およびインサイチュで膨潤性ポリマーを形成することを含む、[30]記載の方法。
[36]該組成物が、1つ以上の水溶性モノマー前駆体を含む水溶液である、[35]記載の方法。
[37]該溶液が、アクリレート、アクリルアミドおよびN,N-メチレンビスアクリルアミドを含む、[36]記載の方法。
[38]膨潤性材料がポリアクリレートである、[30]または[35]記載の方法。
[39]該膨潤性材料が、試料中に浸透化される、[30]または[35]記載の方法。
[40]該試料が、膨潤性材料に固定される、[39]記載の方法。
[41]水を添加することにより、該試料が全方向に等しく膨潤し、拡大された試料を生じる、[30]記載の方法。
[42]水の添加により、埋め込まれた試料が、元の大きさの4x〜5xまで膨張する、[41]記載の方法。
[43]該試料に1つ以上のモノマー前駆体を浸透化させる前に、該試料を界面活性剤で処理する、[35]記載の方法。
[44]該試料を膨潤させる前に、該試料を酵素消化に供し、内部にある内因性生物学的分子を破壊する、[30]記載の方法。
[45]該消化が、標識をそのままで、かつ膨潤性材料に固定させたままにする、[44]記載の方法。
[46]目的の試料自体を物理的に膨張させることに基づく、古典的な顕微鏡法回析限界よりも良好な解像度での目的の試料の光学的画像化のための方法であって、目的の試料を膨潤性材料に埋め込む工程、該材料を膨潤させる工程、および顕微鏡下で該試料を検分する工程を含む、方法。
[47]該試料が標識される、[46]記載の方法。
[48]該試料が、免疫蛍光染色、免疫組織化学的染色または免疫細胞化学的染色により標識される、[47]記載の方法。
[49]標識された試料が、膨潤性材料に固定される、[47]記載の方法。
[50]該標識が、膨潤性材料に結合される、[49]記載の方法。
[51]該試料を膨潤性材料に埋め込む工程が、該試料に、膨潤性ポリマーの前駆体を含む組成物を浸透化させること、およびインサイチュで膨潤性ポリマーを形成することを含む、[46]記載の方法。
[52]該組成物が、1つ以上の膨張性モノマー前駆体を含む水溶液である、[51]記載の方法。
[53]該溶液が、アクリレート、アクリルアミドおよびN,N-メチレンビスアクリルアミドを含む、[52]記載の方法。
[54]膨潤性材料がポリアクリレートである、[46]または[51]記載の方法。
[55]膨潤性材料が、試料中に浸透化される、[46]または[51]記載の方法。
[56]該試料が、膨潤性材料に固定される、[55]記載の方法。
[57]水を添加することにより、試料を全方向に等しく膨潤させ、拡大された試料を生じる、[46]記載の方法。
[58]水の添加により、埋め込まれた試料が、元の大きさの4x〜5xまで膨張する、[57]記載の方法。
[59]該試料に1つ以上のモノマー前駆体を浸透化させる前に、該試料を界面活性剤で処理する、[51]記載の方法。
[60]該試料を膨潤させる前に、該試料を酵素消化に供し、内部にある内因性生物学的分子を破壊する、[46]記載の方法。
[61]該消化が、標識をそのままで、かつ膨潤性材料に固定させたままにする、[60]記載の方法。
[62]膨潤性材料に埋め込まれた目的の試料を含む、顕微鏡法のための膨張可能な(expandable)目的の試料。
[63]該試料が標識される、[62]記載の試料。
[64]該試料が、免疫蛍光染色、免疫組織化学的染色または免疫細胞化学的染色により標識される、[63]記載の試料。
[65]標識された試料が、膨潤性材料に固定される、[63]記載の試料。
[66]該標識が、膨潤性材料に結合される、[65]記載の試料。
[67]該試料を膨潤性材料に埋め込む工程が、該試料に、膨潤性ポリマーの前駆体を含む組成物を浸透化させること、およびインサイチュで膨潤性ポリマーを形成することを含む、[62]記載の試料。
[68]該組成物が、1つ以上の膨張性モノマー前駆体を含む水溶液である、[67]記載の試料。
[69]該溶液が、アクリレート、アクリルアミドおよびN,N-メチレンビスアクリルアミドを含む、[68]記載の試料。
[70]膨潤性材料がポリアクリレートである、[62]または[67]記載の試料。
[71]該膨潤性材料が、試料中に浸透化される、[62]または[67]記載の試料。
[72]該試料が、膨潤性材料に固定される、[71]記載の試料。
[73]該試料に1つ以上のモノマー前駆体を浸透化させる前に、該試料を界面活性剤で処理する、[67]記載の試料。
[74][1]、[2]、[17]または[18]のいずれか一項に記載の方法により作製される、膨張可能な目的の試料。
[75]膨潤性材料に埋め込まれた目的の試料を含み、3次元の全方向に等しい試料の膨張を特徴とする、拡大された複合体。
Claims (1)
- 顕微鏡法のための目的の試料を拡大させるための方法であって、目的の試料を膨潤性(swellable)材料に埋め込む工程、および該材料を膨潤させる工程を含む、方法。
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WO2024071918A1 (ko) * | 2022-09-26 | 2024-04-04 | 연세대학교 산학협력단 | 고배율의 세포 확대 분석을 위한 세포 필름, 이의 제조용 조성물 및 제조 방법 |
Families Citing this family (55)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2014531908A (ja) | 2011-10-14 | 2014-12-04 | プレジデント アンド フェローズ オブ ハーバード カレッジ | 構造アッセンブリによる配列決定 |
US11021737B2 (en) | 2011-12-22 | 2021-06-01 | President And Fellows Of Harvard College | Compositions and methods for analyte detection |
EP4108782B1 (en) | 2011-12-22 | 2023-06-07 | President and Fellows of Harvard College | Compositions and methods for analyte detection |
US9914967B2 (en) | 2012-06-05 | 2018-03-13 | President And Fellows Of Harvard College | Spatial sequencing of nucleic acids using DNA origami probes |
US10138509B2 (en) | 2013-03-12 | 2018-11-27 | President And Fellows Of Harvard College | Method for generating a three-dimensional nucleic acid containing matrix |
KR102512979B1 (ko) | 2013-06-04 | 2023-03-22 | 프레지던트 앤드 펠로우즈 오브 하바드 칼리지 | Rna-가이드된 전사 조절 |
EP4234716A3 (en) | 2013-06-25 | 2023-12-06 | Prognosys Biosciences, Inc. | Methods for determining spatial patterns of biological targets in a sample |
WO2016018963A1 (en) | 2014-07-30 | 2016-02-04 | President And Fellows Of Harvard College | Probe library construction |
CA2982146A1 (en) | 2015-04-10 | 2016-10-13 | Spatial Transcriptomics Ab | Spatially distinguished, multiplex nucleic acid analysis of biological specimens |
US10059990B2 (en) | 2015-04-14 | 2018-08-28 | Massachusetts Institute Of Technology | In situ nucleic acid sequencing of expanded biological samples |
US10526649B2 (en) | 2015-04-14 | 2020-01-07 | Massachusetts Institute Of Technology | Augmenting in situ nucleic acid sequencing of expanded biological samples with in vitro sequence information |
US11408890B2 (en) | 2015-04-14 | 2022-08-09 | Massachusetts Institute Of Technology | Iterative expansion microscopy |
WO2017027368A1 (en) * | 2015-08-07 | 2017-02-16 | Massachusetts Institute Of Technology | Protein retention expansion microscopy |
EP3332029B1 (en) * | 2015-08-07 | 2021-10-06 | Massachusetts Institute of Technology | Nanoscale imaging of proteins and nucleic acids via expansion microscopy |
MX2018005611A (es) | 2015-11-03 | 2018-11-09 | Harvard College | Metodo y aparato para la formacion de imagenes volumetricas de una matriz tridimensional que contiene acido nucleico. |
CA3015363A1 (en) * | 2016-02-25 | 2017-08-31 | Massachusetts Institute Of Technology | Methods for expanding clinical tissue specimens |
US20170276578A1 (en) * | 2016-03-22 | 2017-09-28 | University Of Washington | Expansion microscopy methods and kits |
CN116200465A (zh) | 2016-04-25 | 2023-06-02 | 哈佛学院董事及会员团体 | 用于原位分子检测的杂交链反应方法 |
US11397140B2 (en) | 2016-04-29 | 2022-07-26 | Massachusetts Institute Of Technology | Methods for reversible and tunable tissue magnification |
WO2017222453A1 (en) | 2016-06-21 | 2017-12-28 | Hauling Thomas | Nucleic acid sequencing |
CN109923216B (zh) | 2016-08-31 | 2024-08-02 | 哈佛学院董事及会员团体 | 将生物分子的检测组合到使用荧光原位测序的单个试验的方法 |
CN109983125B (zh) | 2016-08-31 | 2024-06-04 | 哈佛学院董事及会员团体 | 生成用于通过荧光原位测序检测的核酸序列文库的方法 |
EP4339956A3 (en) * | 2016-11-08 | 2024-06-19 | President and Fellows of Harvard College | Multiplexed imaging using merfish, expansion microscopy, and related technologies |
US10995361B2 (en) | 2017-01-23 | 2021-05-04 | 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 |
CN106959240B (zh) * | 2017-03-28 | 2019-09-24 | 华中科技大学 | 一种反复膨胀过程实现大块组织体的膨胀透明方法 |
US11788123B2 (en) | 2017-05-26 | 2023-10-17 | President And Fellows Of Harvard College | Systems and methods for high-throughput image-based screening |
US11719606B2 (en) * | 2017-07-06 | 2023-08-08 | University Public Corporation Osaka | Method and reagent for clearing biological tissue |
WO2019023214A1 (en) | 2017-07-25 | 2019-01-31 | Massachusetts Institute Of Technology | ATAC IN SITU SEQUENCING |
CA3078158A1 (en) | 2017-10-06 | 2019-04-11 | Cartana Ab | Rna templated ligation |
WO2019075091A1 (en) | 2017-10-11 | 2019-04-18 | Expansion Technologies | MULTIPLEXED IN SITU HYBRIDIZATION OF TISSUE SECTIONS FOR SPATIALLY RESOLVED TRANSCRIPTOMIC WITH EXPANSION MICROSCOPY |
US11479811B2 (en) | 2017-11-21 | 2022-10-25 | Expansion Technologies | Expansion microscopy compatible and multiplexed in situ hybridization of formalin fixed paraffin embedded tissue sections for spatially resolved transcriptomics |
WO2019156957A1 (en) | 2018-02-06 | 2019-08-15 | Massachusetts Institute Of Technology | Swellable and structurally homogenous hydrogels and methods of use thereof |
US10915729B2 (en) | 2018-02-20 | 2021-02-09 | The Regents Of The University Of Michigan | Three-dimensional cell and tissue image analysis for cellular and sub-cellular morphological modeling and classification |
JP7082364B2 (ja) * | 2018-03-02 | 2022-06-08 | 日本電信電話株式会社 | フィルム状ソフトマテリアル、蛍光フィルム基材及びフィルム状ソフトマテリアルの製造方法、並びに応力計測方法 |
WO2019209743A1 (en) * | 2018-04-25 | 2019-10-31 | The Regents Of The University Of California | Histology-grade three-dimensional imaging of tissue using microscopy with ultraviolet surface excitation |
US11414701B2 (en) | 2018-05-24 | 2022-08-16 | The Broad Institute, Inc. | Multimodal readouts for quantifying and sequencing nucleic acids in single cells |
WO2020013833A1 (en) * | 2018-07-13 | 2020-01-16 | Massachusetts Institute Of Technology | Dimethylacrylamide (dmaa) hydrogel for expansion microscopy (exm) |
SG11202101934SA (en) | 2018-07-30 | 2021-03-30 | Readcoor Llc | Methods and systems for sample processing or analysis |
US11549135B2 (en) | 2018-09-14 | 2023-01-10 | The Broad Institute, Inc. | Oligonucleotide-coupled antibodies for single cell or single complex protein measurements |
KR102368585B1 (ko) * | 2018-09-28 | 2022-03-03 | 서울대학교산학협력단 | 온도에 따라 가역적 팽창/초기화가 가능한 하이드로젤을 이용한 생체조직 이미징 |
CA3130889A1 (en) * | 2019-02-22 | 2020-08-27 | Massachusetts Institute Of Technology | Iterative direct expansion microscopy |
CN110006862B (zh) * | 2019-03-29 | 2020-09-08 | 华中科技大学 | 膨胀切削显微成像方法及适用于该方法的超吸水水凝胶 |
CA3139791A1 (en) | 2019-05-31 | 2020-12-03 | 10X Genomics, Inc. | Method of detecting target nucleic acid molecules |
US11802822B2 (en) | 2019-12-05 | 2023-10-31 | Massachusetts Institute Of Technology | Multiplexed expansion (MultiExM) pathology |
EP4153606A2 (en) | 2020-07-13 | 2023-03-29 | Singular Genomics Systems, Inc. | Methods of sequencing complementary polynucleotides |
WO2022032195A2 (en) | 2020-08-06 | 2022-02-10 | Singular Genomics Systems, Inc. | Spatial sequencing |
EP4121561A4 (en) | 2020-08-06 | 2024-04-17 | Singular Genomics Systems, Inc. | METHODS FOR IN SITU TRANSCRIPTOMICS AND PROTEOMICS |
US12071667B2 (en) | 2020-11-04 | 2024-08-27 | 10X Genomics, Inc. | Sequence analysis using meta-stable nucleic acid molecules |
US11333588B1 (en) | 2020-12-07 | 2022-05-17 | Nebulum Technologies Co., Ltd. | Matrix-assisted methods and compositions to prepare biological samples for super-resolution imaging |
CN112881671A (zh) * | 2021-01-11 | 2021-06-01 | 东南大学 | 一种基于高密度荧光标记法的细胞微管膨胀显微成像方法 |
US12060603B2 (en) | 2021-01-19 | 2024-08-13 | 10X Genomics, Inc. | Methods for internally controlled in situ assays using padlock probes |
WO2023220300A1 (en) | 2022-05-11 | 2023-11-16 | 10X Genomics, Inc. | Compositions and methods for in situ sequencing |
DE102022112065B3 (de) | 2022-05-13 | 2023-08-24 | Abberior Instruments Gmbh | Verfahren, vorrichtung und computerprogramm zur mikroskopischen probenabbildung |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2006036957A (ja) * | 2004-07-28 | 2006-02-09 | Oita Univ | 高親水性高分子による組織包埋方法 |
JP2008286694A (ja) * | 2007-05-18 | 2008-11-27 | Shinichiro Isobe | 生体標本の作製方法 |
JP2014005231A (ja) * | 2012-06-22 | 2014-01-16 | Institute Of Physical & Chemical Research | 生物材料を透明化する方法および生物材料用透明化処理キット |
WO2014025392A1 (en) * | 2012-08-09 | 2014-02-13 | The Board Of Trustees Of The Leland Stanford Junior University | Methods and compositions for preparing biological specimens for microscopic analysis |
Family Cites Families (15)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6107081A (en) * | 1999-02-05 | 2000-08-22 | The United States Of America As Represented By The Administrator Of The National Aeronautics And Space Administration | Uni-directional cell stretching device |
WO2000066265A2 (en) * | 1999-04-27 | 2000-11-09 | Ciphergen Biosystems, Inc. | Probes for a gas phase ion spectrometer |
US6287870B1 (en) * | 1999-08-20 | 2001-09-11 | Robert A. Levine | Method and assembly for separating formed constituents from a liquid constituent in a complex biologic fluid sample |
US6878384B2 (en) * | 2001-03-13 | 2005-04-12 | Microvention, Inc. | Hydrogels that undergo volumetric expansion in response to changes in their environment and their methods of manufacture and use |
JP3812829B2 (ja) * | 2002-08-22 | 2006-08-23 | 独立行政法人科学技術振興機構 | 組織の包埋剤、樹脂包埋体及び樹脂包埋体の製造方法 |
JP2005291759A (ja) | 2004-03-31 | 2005-10-20 | Michimasa Kishimoto | 二次元画像による病症診断システム |
ES2348126T3 (es) * | 2007-02-01 | 2010-11-30 | Immundiagnostik Ag | Determinación directa de la vitamina d en suero o plasma. |
EP2195413A4 (en) | 2007-08-30 | 2014-01-01 | Harvard College | MULTI-WELL CULTURE PLATE HAVING A HIGH COMPATIBILITY SURFACE |
US20100041128A1 (en) | 2008-01-08 | 2010-02-18 | Medtrain Technologies, Llc | Microfluidic Device for Application of Shear Stress and Tensile Strain |
WO2009099552A2 (en) | 2008-01-30 | 2009-08-13 | Corning Incorporated | Cell culture article and screening |
JP4956839B2 (ja) * | 2008-02-13 | 2012-06-20 | 国立大学法人 大分大学 | 高親水性高分子モノマー水溶液による組織包埋方法 |
US20090241681A1 (en) * | 2008-03-27 | 2009-10-01 | Andrew Machauf | Hydrogel-based mems biosensor |
RU2010151919A (ru) | 2008-05-20 | 2012-06-27 | Зе Реджентс Оф Зе Юниверсити Оф Калифорния (Us) | Анализ ex vivo клеток с целью детектирования болезненного состояния и выбора и мониторинга терапевтического агента |
WO2011111876A1 (en) * | 2010-03-12 | 2011-09-15 | Riken | Clearing reagent for biological material, and use thereof |
US20140087412A1 (en) | 2011-04-20 | 2014-03-27 | 4Dx Pty Ltd | Method and Device for Application of Fluid Forces to Cells |
-
2015
- 2015-02-20 US US14/627,310 patent/US10309879B2/en active Active
- 2015-02-20 WO PCT/US2015/016788 patent/WO2015127183A2/en active Application Filing
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- 2015-02-20 JP JP2016553349A patent/JP6456969B2/ja active Active
-
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- 2018-10-26 JP JP2018202178A patent/JP2019056708A/ja active Pending
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2006036957A (ja) * | 2004-07-28 | 2006-02-09 | Oita Univ | 高親水性高分子による組織包埋方法 |
JP2008286694A (ja) * | 2007-05-18 | 2008-11-27 | Shinichiro Isobe | 生体標本の作製方法 |
JP2014005231A (ja) * | 2012-06-22 | 2014-01-16 | Institute Of Physical & Chemical Research | 生物材料を透明化する方法および生物材料用透明化処理キット |
WO2014025392A1 (en) * | 2012-08-09 | 2014-02-13 | The Board Of Trustees Of The Leland Stanford Junior University | Methods and compositions for preparing biological specimens for microscopic analysis |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2024071918A1 (ko) * | 2022-09-26 | 2024-04-04 | 연세대학교 산학협력단 | 고배율의 세포 확대 분석을 위한 세포 필름, 이의 제조용 조성물 및 제조 방법 |
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