JP6960696B2 - 磁性−光学複合ナノ構造体 - Google Patents
磁性−光学複合ナノ構造体 Download PDFInfo
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- JP6960696B2 JP6960696B2 JP2020136255A JP2020136255A JP6960696B2 JP 6960696 B2 JP6960696 B2 JP 6960696B2 JP 2020136255 A JP2020136255 A JP 2020136255A JP 2020136255 A JP2020136255 A JP 2020136255A JP 6960696 B2 JP6960696 B2 JP 6960696B2
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Description
前記第1のコア−シェルナノ粒子に金ナノ粒子を付着させて金ナノ粒子が付着したコア−シェルナノ粒子を製造する段階と、
前記金ナノ粒子が付着したコア−シェルナノ粒子の前記金ナノ粒子を1次成長させる段階と、
前記1次成長した金ナノ粒子上にラマン分子を機能化させてラマン分子が機能化されたコア−シェルナノ粒子を製造する段階と、
前記ラマン分子が機能化されたコア−シェルナノ粒子のラマン分子が機能化された金ナノ粒子に金、銀、又は金−銀合金シェルを形成して第2のコア−シェルナノ粒子を形成する段階と、を含む磁性−光学複合ナノ構造体の製造方法を提供する。
磁性ナノ粒子コア及びシリカシェルを含む第1のコア−シェルナノ粒子と、
金ナノ粒子コア及び金、銀又は金と銀の合金であるシェルを含む第2のコア−シェルナノ粒子と、を含み、
前記第1のコア−シェルナノ粒子のシェルの表面に形成された官能基に第2のコア−シェルナノ粒子が結合されており、
前記金ナノ粒子コアは、ラマン分子として機能化されている磁性−光学複合ナノ構造体を提供する。
前記機能化された磁性−光学複合ナノ構造体を一つ以上の分析物を含む試料に露出させる段階と、
ラマン分光法を用いて磁性−光学複合ナノ構造体に結合された分析物を確認する段階と、を含む、分析物を検出、画像化又は分離する方法を提供する。
(B)前記第1のコア−シェルナノ粒子に金ナノ粒子を付着させて金ナノ粒子が付着したコア−シェルナノ粒子を製造する段階(以下、金ナノ粒子が付着したコア−シェルナノ粒子製造段階)と、
(C)前記金ナノ粒子が付着したコア−シェルナノ粒子の前記金ナノ粒子を1次成長させる段階(以下、1次成長段階)と、
(D)前記1次成長した金ナノ粒子上にラマン分子を機能化させてラマン分子が機能化されたコア−シェルナノ粒子を製造する段階(以下、ラマン分子が機能化されたコア−シェルナノ粒子製造段階)と、
(E)前記ラマン分子が機能化されたコア−シェルナノ粒子のラマン分子が機能化された金ナノ粒子に金、銀、又は金−銀合金シェルを形成して第2のコア−シェルナノ粒子を形成する段階(以下、2次成長段階)と、を含んでもよい。
金ナノ粒子コア及び金、銀又は金と銀の合金であるシェルを含む第2のコア−シェルナノ粒子を含んでもよい。
前記機能化された磁性−光学複合ナノ構造体を一つ以上の分析物を含む試料に露出させる段階と、
ラマン分光法を用いて磁性−光学複合ナノ構造体に結合された分析物を確認する段階と、を含む、分析物を検出、分離、又は画像化する方法に関する。
実施例1.磁性−光学ナノ複合構造体の製造
1.酸化鉄(磁鉄鉱、magnetite、Fe3O4)ナノ粒子の合成
塩化金水和物(gold chloride trihydrate、HAuCl4・3H2O)は、金イオン前駆体として、水素化ホウ素ナトリウム(sodium borohydride、NaBH4)は、還元剤として、クエン酸ナトリウム(trisodium citrate)は、安定化剤として、H2Oは、溶媒として使用した。
酸化鉄ナノ粒子にストーバー(Stober)法を用いてシリカ(silica)シェル(shell)を形成した。
酸化鉄ナノ粒子−シリカコア−シェルナノ粒子上の金ナノ粒子を1次成長させるため、PVPは、安定化剤として、四塩化金酸水和物(HAuCl4・3H2O)は、金イオン前駆体として、アスコルビン酸(L−ascorbic acid)は、還元剤として、H2Oは、溶媒として使用した。
ラマン分子で機能化していないことを除いては、実施例1と同様の方法で磁性−光学ナノ複合構造体を製造した。
図1の(a)は、本発明による磁性−光学複合ナノ構造体の製造過程(段階)模式図であり、(b)は、前記模式図において、それぞれの段階の透過電子顕微鏡(Transmission Electron Microscopy、TEM)写真であり、(c)は、(S3)で製造時の透過電子顕微鏡写真、及びFe、Si、Au、S(ラマン分子のサイオール基(−SH))及びAgのエネルギー分散型分光分析法(Energy Dispersive X−ray Spectroscopy、EDX)写真を示す。
実施例で製造された磁性−光学複合ナノ構造体をカバーグラス(cover glass)上に乾燥させて、単一ナノ構造体に対する表面増強ラマン散乱(SERS)信号の強度を共焦点ラマン顕微鏡(Confocal Raman Microscopy)を用いて測定した。このとき、100倍対物レンズを使用し、レーザー波長は、633nmと785nmを使用した。レーザー波長633nmの照射時のレーザーパワーは、1mW、獲得時間(acquisition time)は、1秒測定した。レーザー波長785nmの照射時、レーザーパワーは、4mW、獲得時間(acquisition time)は、5秒測定した。
実験例2で製造されたcyclo(−RGDyK)で機能化された複合ナノ構造体溶液0.05mLをU87MG細胞が130,000個分散されているPBS溶液と30分間培養(incubation)し、PBS溶液で磁性分離を用いて洗浄した後、1mL PBS溶液に再分散した。
実験例2で製造されたcyclo(−RGDyK)で機能化された複合ナノ構造体溶液0.050mLをU87MG細胞が130,000個のPBS溶液と30分間培養し、PBS溶液で磁性分離を用いて洗浄した。そして、1mL PBS溶液に再分散して磁性分離された細胞の数を計算した。
(S3)に該当する複合ナノ構造体のカルボキシル基の表面機能化のため、SH−PEG−COOH(molecular weight5000、MW5000)、H2Oを溶媒として、sodium dodecyl sulfate(SDS)を安定化剤として使用した。
Claims (17)
- 磁性ナノ粒子にセラミックシェルを形成して第1のコア−シェルナノ粒子を製造する段階と、
前記第1のコア−シェルナノ粒子に金ナノ粒子を付着させて金ナノ粒子が付着したコア−シェルナノ粒子を製造する段階と、
前記金ナノ粒子が付着したコア−シェルナノ粒子の前記金ナノ粒子を1次成長させる段階と、
前記1次成長した金ナノ粒子上にラマン分子を機能化させてラマン分子が機能化されたコア−シェルナノ粒子を製造する段階と、
前記ラマン分子が機能化されたコア−シェルナノ粒子のラマン分子が機能化された金ナノ粒子に金、銀、又は金−銀合金シェルを形成して第2のコア−シェルナノ粒子を形成する段階と、を含む、
磁性−光学複合ナノ構造体の製造方法。 - 磁性ナノ粒子は、FeO、Fe2O3、Fe3O4、CoFe2O4、NiFe2O4、MnFe2O4、TiO2、ZrO2、CeO2、Al2O3、及びMgOからなるグループから選ばれた一つ以上の金属酸化物ナノ粒子である、請求項1に記載の磁性−光学複合ナノ構造体の製造方法。
- 磁性ナノ粒子の平均粒径は、10〜500nmである、請求項1に記載の磁性−光学複合ナノ構造体の製造方法。
- セラミックは、シリカ、チタニア、ジルコニア、アルミナ、及びゼオライトからなるグループから選ばれた一つ以上を含むものである、請求項1に記載の磁性−光学複合ナノ構造体の製造方法。
- 金ナノ粒子が付着したコア−シェルナノ粒子を製造する段階は、
第1のコア−シェルナノ粒子のシェルの表面に官能基を導入する段階と、
前記官能基に金ナノ粒子シードを結合させる段階と、を含むものである、請求項1に記載の磁性−光学複合ナノ構造体の製造方法。 - 官能基は、アミン基(−NH)、サイオール基(−SH)、カルボキシル基(−COOH)、ヒドロキシル基(−OH)、及びドーパミンからなるグループから選ばれた一つ以上を含むものである、請求項5に記載の磁性−光学複合ナノ構造体の製造方法。
- 金ナノ粒子を1次成長させる段階において、前記成長した金ナノ粒子の平均粒径は、5〜50nmである、請求項1に記載の磁性−光学複合ナノ構造体の製造方法。
- ラマン分子が機能化されたコア−シェルナノ粒子を製造する段階において、ラマン分子は、1,4−BDT(1,4−ベンゼンジチオール)、FAM(フルオレセイン)、ダブシル(Dabcyl)、TRIT(テトラメチルローダミンイソチオール)、NBD(7−ニトロベンズ2−1,3−ダイヤゾール)、テキサスレッド(Texas Red)染料、フタル酸、テレフタル酸、イソフタル酸、クレシルファストバイオレット、クレシルブルーバイオレット、ブリリアント(brilliant)クレシルブルー、パラ−アミノ安息香酸、エリスロシン、ビオチン、ジゴキシゲニン(digoxigenin)、5−カルボキシ−4’,5’−ジクロロ−2’,7’−ジメトキシ、5−カルボキシ−2’,4’,5’,7’−テトラクロロフルオレセイン、5−カルボキシフルオレセイン、5−カルボキシローダミン、6−カルボキシローダミン、6−カルボキシテトラメチルアミノフタロシアニン、アゾメチン、キサンチン、スクシニルフルオレセイン、アミノアクリジン、量子点、カーボンナノチューブ、炭素同素体、シアン化物、チオール、クロリン、臭素、メチル、リン、硫黄及びシアニン染料(Cy3、Cy3.5、Cy5)及びローダミンからなるグループから選ばれた一つ以上を含むものである、請求項1に記載の磁性−光学複合ナノ構造体の製造方法。
- 金、銀、又は金−銀合金シェルを形成する段階において、金イオン前駆体溶液及び銀イオン前駆体溶液の金イオン及び銀イオンの濃度は、それぞれ0.01mM〜1Mであり、
前記金イオン前駆体溶液及び銀イオン前駆体溶液の重量比は、1:0〜0:1である、請求項1に記載の磁性−光学複合ナノ構造体の製造方法。 - 金イオン前駆体溶液のみを使用するか、又は金イオン及び銀イオンの混合含量に対して金イオンの含量が50モル%以上である金イオン前駆体溶液及び銀イオン前駆体溶液の混合溶液を使用すると、シェルは、エピタキシャル成長パターンを示すものである、請求項9に記載の磁性−光学複合ナノ構造体の製造方法。
- エピタキシャルに成長した第2のコア−シェルナノ粒子のサイズは、5nm〜100nmである、請求項10に記載の磁性−光学複合ナノ構造体の製造方法。
- 銀イオン前駆体溶液のみを使用するか、又は金イオン及び銀イオンの混合含量に対して銀イオンの含量が50モル%を超える金イオン前駆体溶液及び銀イオン前駆体溶液の混合溶液を使用すると、シェルは、銀アイランド(island)に成長するものである、請求項9に記載の磁性−光学複合ナノ構造体の製造方法。
- 銀アイランドに成長した第2のコア−シェルナノ粒子のサイズは、5nm〜500nmである、請求項12に記載の磁性−光学複合ナノ構造体の製造方法。
- 磁性ナノ粒子コア及びセラミックシェルを含む第1のコア−シェルナノ粒子と、
金ナノ粒子コア及び金、銀又は金と銀の合金であるシェルを含む第2のコア−シェルナノ粒子を含み、
前記第1のコア−シェルナノ粒子のシェルの表面に形成された官能基に第2のコア−シェルナノ粒子が結合されており、
前記金ナノ粒子コアは、ラマン分子で機能化されている、磁性−光学複合ナノ構造体。 - 第2のコア−シェルナノ粒子は、前記第1のコア−シェルナノ粒子の表面よりも内部に位置しているものである、請求項14に記載の磁性−光学複合ナノ構造体。
- 請求項14に記載の磁性−光学複合ナノ構造体を含む分析物検出用キット、分子診断チップ又は画像診断用組成物。
- 請求項14に記載の磁性−光学複合ナノ構造体の表面に検出しようとする分析物と結合できるバイオ分子を機能化する段階と、
前記機能化された磁性−光学複合ナノ構造体を一つ以上の分析物を含む試料に露出させる段階と、
ラマン分光法を用いて磁性−光学複合ナノ構造体に結合された分析物を確認する段階と、を含む
分析物を検出、分離又は画像化する方法。
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