JP2006514708A - シリカ系蛍光ナノ粒子 - Google Patents

シリカ系蛍光ナノ粒子 Download PDF

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JP2006514708A
JP2006514708A JP2004568576A JP2004568576A JP2006514708A JP 2006514708 A JP2006514708 A JP 2006514708A JP 2004568576 A JP2004568576 A JP 2004568576A JP 2004568576 A JP2004568576 A JP 2004568576A JP 2006514708 A JP2006514708 A JP 2006514708A
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ウィーズナー，ウルリッヒ
オウ，フーイスウェング
ラーソン，ダニエル，アール．
ウェッブ，ワット，ダブリュー．
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Abstract

【解決すべき課題】
新規なシリカ系の蛍光ナノ粒子を発明すること。
【解決手段】
本発明は一般的に、蛍光ナノ粒子に関するものであり、より具体的には、有機色素が共有結合している３０ｎｍより小さいシリカ系の蛍光ナノ粒子に関する。本発明は、発蛍光団中心コア、及びシリカ殻を含む蛍光単分散シリカナノ粒子を提供する。このナノ粒子の放射特性は、コアの化学的性質（組成）とシリカ殻の存在に依存する。本発明は概して、３０ｎｍまで又はそれ以下の大きさのシリカ粒子内に包埋された色素分子の光物理的特性の制御を提供する。この制御はシリカの化学的性質と粒子構造をナノメートル規模で変えることにより達成されるものであり、その結果、遊離色素と比べて顕著な輝度の向上が得られる。

Description

本発明は一般的には、ナノ粒子に関し、そしてより具体的には、有機色素に共有付着している３０ｎｍより小さいシリカ系のナノ粒子に関する。
本出願は、参照によりインコーポレートされる２００２年11月２６日に出願された非仮出願米国特許出願番号第１０／３０６６１４に対する優先権を主張するものである。
蛍光ナノ粒子は、例えば生体画像化、センサー技術、マイクロアレイ、及び光コンピューティングなど、バイオテクノロジーと情報の適用の指標及び光子源として、とてつもなく有望である。これらの適用には、生体高分子に特異的に結合できるか又は高次構造に配置できる、サイズ管理された、単分散の、明るいナノ粒子が要求される。
最近、一連の要求の厳しい生物学的適用及び分析適用における使用に向けた、高蛍光性水溶性ナノ粒子を合成するための一般的なアプローチが二つ出現した。最初のアプローチでは、ナノ粒子状物質自体が蛍光性（半導体ナノ結晶、または金属ナノ結晶など）である。二つ目のアプローチでは、蛍光ナノ粒子は有機色素分子の取り込みに基づくものである。有機色素分子の多様性が大きく、これら色素分子の局所環境に対する鋭敏な感受性を考えると、後者のアプローチでは、精密に制御された広範囲の蛍光特性を持つナノ粒子を開発できる可能性が高くなる。
ステーバーらの研究に基づき、色素を包埋したシリカナノ粒子が、様々なサイズ、色、構造で合成されてきた。光物理的性質の全ての先行報告では、シリカ粒子内部で共有結合している色素は遊離色素と比較して消光されることが観察されている。しかしながら、その内部で色素が拡散により非共有的に付着及び負荷されているポリ（オルガノシロキサン）マイクロゲルの場合は、蛍光効率の僅かな向上が観察される。他の物質、例えばポリスチレンミクロスフェアについては、包埋色素の量子効率は遊離色素と同じか又は遊離色素よりも低い。蛍光の消光は、通常はシリカ媒体中へのシリカ粒子内エネルギー転移又は非放射減衰のいずれかに起因するものである。これらの経路はいずれも、粒子内の局所的色素環境に影響される可能性があり、このことは粒子内部における構造の正確な制御が消光を回復させるか、又はさらに蛍光増強をもたらすかもしれない。
記載なし 記載なし
有機色素が共有結合した３０ｎｍより小さい高蛍光性ナノ粒子がやはり必要である。
発明の概要
本発明は、シリカ粒子内の構造が構成色素の放射特性に対する顕著な且つ制御された効果を有する、あるタイプのシリカナノ粒子に関する。
本発明は、発蛍光団中心コアとシリカ殻を含む蛍光単分散シリカナノ粒子を提供する。該ナノ粒子の放射特性は、該コアの化学的性質（組成）とシリカ殻の存在に依存する。本発明の一側面では、コア−殻構造は蛍光量子効率の向上をもたらす。本発明は一般的に、３０ｎｍ以下のサイズのシリカ粒子に被包された色素分子の光物理的性質の制御を提供する。この制御は、シリカの化学的性質とナノメートルのサイズ規模の粒子構造における変化により得られ、遊離色素と比べてはっきりした輝度の増強をもたらす。
本発明の別の一側面では、コア−殻構造はナノ粒子の光安定性を制御することを可能にする。
本発明によりもたらされる量子効率の向上は放射速度の増加と非放射速度の減少の両方によるものであり、後者の影響は構造間で最も変わりやすい。非放射速度の変化は、粒子内色素の回転運動性の差異とよく相関する。
本発明の一つの実施態様では、蛍光単分散ナノ粒子にはコア−殻構造が含まれ、そのコアはシリカ殻で囲まれた密集コア（compact core）を含む。本発明の別の一実施態様では、蛍光単分散ナノ粒子にはコア−殻構造が含まれ、コアはシリカ殻で囲まれた膨張コア（expanded core）を含む。本発明の別の実施態様では、蛍光単分散ナノ粒子にはコア−核構造が含まれ、この構造内ではコアは色素がシリカ殻に囲まれた範囲内で低密度に包埋された均質粒子を含む。本発明のまた別の実施態様では、蛍光単分散均質ナノ粒子はシリカ殻に囲まれていない。
本発明は、蛍光化合物とオルガノシラン化合物を混合して色素前駆体を形成し、その結果得られた色素前駆体を水溶液と混合して密集蛍光コアを形成し、その結果得られた密集コアとシリカ前駆体を混合して密集コア上にシリカ殻を形成することによって蛍光単分散ナノ粒子を得る、密集コア構造を有する蛍光単分散ナノ粒子の製造法を提供する。
本発明はまた、蛍光化合物とオルガノシラン化合物を混合して色素前駆体を形成し、その結果得られた色素前駆体をシリカ前駆体と共縮合して膨張コアを形成し、その結果得られた膨張コアとシリカ前駆体を混合して膨張コア上にシリカ殻を形成することによって蛍光単分散ナノ粒子を得る、膨張コア構造を有する蛍光単分散ナノ粒子の製造法を提供する。
本発明は、蛍光化合物とオルガノシラン化合物を混合して色素前駆体を形成し、その結果得られた色素前駆体をシリカ前駆体と共縮合して均質な蛍光単分散ナノ粒子を形成することにより均質（homogenous）な蛍光単分散ナノ粒子の製造法を提供する。
本発明のこれらの特性及びその他の特性は、以下の本発明の説明に記述する。
本発明の蛍光ナノ粒子には、蛍光シラン化合物を含むコア、及びコア上のシリカ殻が含まれる。ナノ粒子のコアは、例えば反応性蛍光化合物と共反応性有機シラン化合物の反応生成物を含むことができ、殻は、例えばシリカ形成化合物の反応生成物を含むことができる。シリカ形成化合物は、例えば１層以上（例えば１層〜２０層など）のシリカ層、並びに、殻層の厚み、コア厚み若しくはコア直径に対する殻厚みの比率、コアのシリカ殻表面被覆度、シリカ殻の多孔度と収容力など様々な所望の殻特性を製造することができる。
蛍光単分散コア−殻ナノ粒子の合成は、二段階の工程に基づく、第一に、有機色素分子であるテトラメチルローダミン・イソチオシアネート（ＴＲＩＴＣ）を、シリカ前駆体に共有結合させ、色素が豊富なコアを形成するように縮合させる。第二に、シリカゲルモノマーを添加して蛍光コア原料の周囲に密度がより高いシリカネットワークを形成すると、光安定性に有害な溶媒相互作用から遮蔽することができる。最初に色素豊富なコアを形成することにより、ＵＶ−Ｖｉｓ吸光全体スペクトルと発光スペクトルをカバーする様々なタイプの発蛍光団を組み込むことが可能になる。
色素配置の点で異なるコア−殻構造、即ち、密集コア、膨張コア、及び均質の合成に用いられる試薬の量は、正確な構造特性の相関関係を発生させるため、同一に保たれてきた。しかしながら、反応の結果コア−殻構造を形成する試薬の反応順序は変わる。従って、各構造の色素分子周囲の化学的環境は、シリカ媒体密度に関して異なり、そして光物理的な粒子特性に顕著な効果がある有機部分の存在という点で異なる。
密集コア−殻ナノ粒子を合成するため、適量のアンモニア、水、溶媒を含む反応槽に色素前駆体を添加し、一晩反応させた。色素前駆体は、湿度を排除してモル比１：５０のＴＲＩＴＣと３−アミノプロピル・トリエトキシシラン間の付加反応により合成した。色素豊富な密集コアの合成が完了した後、テトラエチルオルソシリケート（ＴＥＯＳ）を引き続き添加して、コアを囲むシリカ殻を成長させた。
膨張コア−殻ナノ粒子の合成は、ＴＥＯＳと上述の色素前駆体を共縮合させ、その混合物を一晩反応させて達成した。膨張コアの合成が完了した後、さらにＴＥＯＳを添加して、コアを囲むシリカ殻を成長させた。
均質ナノ粒子の合成は、全ての試薬、色素前駆体及びＴＥＯＳを同時に共縮合し、その混合物を一晩反応させて達成した。
図２Ａ〜Ｃに示すＦＣＳ曲線と一重指数関数フィットは、水溶液中で各構造に求められる単分散性を示すものである。ＦＣＳ曲線は、さらに二つの独立のパラメータ、即ち、拡散係数と絶対濃度も提供する。拡散係数は流体半径に直接関連し、絶対濃度は粒子輝度の定量を可能にする。
各ナノ粒子とそれぞれの中間体のＦＣＳ曲線を図２Ａ〜Ｃに示す。図２Ａに示すように、密集コア−殻構造のコアの拡散係数（Ｄ＝０．０９８μｍ２／ｍｓ）は、遊離ＴＲＩＴＣ色素の拡散係数（Ｄ＝０．２１μｍ２／ｍｓ）の約半分である。完全なナノ粒子は、図２Ａに示すように約１０倍大きい（Ｄ＝０．０１４μｍ２／ｍｓ）。遊離ＴＲＩＴＣ色素、膨張コア−殻構造、及び完全なナノ粒子間の同様の関係を図２Ｂに示す。膨張コア−殻構造のコアの拡散係数は、遊離色素ＴＲＩＴＣの拡散係数値（Ｄ＝０．２１μｍ２／ｍｓ）と比較して０．０７５μｍ２／ｍｓである。均質ナノ粒子では中間体コア構造はないので、図２Ｃは遊離ＴＲＩＴＣ色素に対する均質ナノ粒子の拡散係数（Ｄ＝０．０１５μｍ２／ｍｓ）を示す。
図２Ｄ〜Ｅは、各構造と合成中間体の輝度を表す。コア中間体で進める合成については、複数の色素が恐らくはコアに存在するという事実にも関わらず、コアは常に遊離色素よりも暗い。完全なナノ粒子の輝度は常に遊離色素及び／又はコア中間体よりも有意に高い。さらに、前駆体物質の粒径と絶対量が同じであるにも関わらず、輝度は構造の違いによって変わる。
本発明の蛍光ナノ粒子には有機色素であるＴＲＩＴＣが含まれる。この色素は、その発光特性における量子効率向上効果を示す。本発明の蛍光ナノ粒子が第一エネルギー源で照射されると、エネルギーの二次発光がナノ粒子で用いられる有機色素のバンドギャップに相当する振動数で発生する。
粒子当たりのＴＲＩＴＣ等価物の数は、ＦＣＳと吸光度測定の組合せにより測定することができる。この組合せでは、図３Ａ〜Ｃに示すように絶対濃度をＦＣＳから測定し、溶液中のＴＲＩＴＣについて相対吸光度を測定する。図３Ａ〜Ｃでは、試料をＦＣＳと吸光度の両方について測定できるよう、希薄溶液（約５０ｎＭ）を用いた。５１４．５ｎｍで４２，１０５Ｍ−１ｃｍ−１という吸光係数を用いて計算した粒子当たりのＴＲＩＴＣ等価物の数を表ＩＩに示す。同様に、ナノ粒子蛍光とナノ粒子吸光度の相対強度は、表ＩＩに示すように遊離ＴＲＩＴＣを超える量子効率の向上を示す。
密集コア−殻及び膨張コア−殻粒子では、粒子当たりのＴＲＩＴＣ等価物の数は８．６で区別が付かない。しかしながら、図３Ａ、３Ｂ及び表ＩＩに示すように、膨張コア−殻ナノ粒子は遊離ＴＲＩＴＣ色素と比べて３倍の量子効率向上を示し、一方密集コア−殻は遊離ＴＲＩＴＣ色素に対して２倍の量子効率向上しか示さない。最大の量子効率向上は均質ナノ粒子で観察され、図３Ｃ及び表ＩＩに示すように、平均で粒子当たり２．３個のＴＲＩＴＣ等価物を有する。
ナノ粒子の化学的性質と構造を通じて光物理的性質を制御できることにより、次世代蛍光プローブの開発と溶液中では得難い蛍光特性の研究の両方が可能になる。
粒子の蛍光寿命を図４Ａに示す。遊離ＴＲＩＴＣを含む各寿命は多重指数関数的であり、平均寿命値を表ＩＩＩにまとめる。ＴＲＩＴＣの寿命（τｆ＝２．１ｎｓ）は、水中での文献値と良く一致している。寿命は、密集コア−殻（１．８ｎｓ）から膨張コア−殻（２．９ｎｓ）へ、均質粒子（３．２ｎｓ）へと増加する（図４Ａ）。密集コア−殻の寿命に最も優先的に貢献するのは高速成分（fast component）であるが、密集コア−殻は遊離色素よりも寿命が短い（図４Ａ）。この寿命データ単独から、色素は密集コア−殻粒子内部で消光すると結論付けてしまうかもしれないが、定常状態の測定はそうではないことを示唆している。蛍光寿命と量子効率の組合せにより、放射速度定数と非放射速度定数の独自の測定が可能になる。表ＩＩＩに正規化形と絶対形の両方を挙げる。放射速度の増加は異なる構造の間で一定であり、遊離のＴＲＩＴＣよりも２．２倍大きい。しかしながら、非放射速度は密集コア−殻ナノ粒子に対する比較的高い値から均質粒子に対する約３倍低い値まで、構造により変化する。
粒子の回転異方性は、ここでも複雑な多重指数関数的な挙動を示し（図４Ｂ、４Ｃ）、我々は現時点では平均回転時間定数（表ＩＩＩ、θ）に限定した。予測どおり、粒子内部の色素はシリカ媒体により妨害され、遊離色素よりも回転時間が長くなる（図４Ｂ）。実際異方性曲線は、クマリン制御とは対照的に、測定の時間目盛上、ゼロまでは減衰しない（遊離ＴＲＩＴＣにも残留異方性がいくらかあることに注意されたい）。しかしながら、緊密に詰め込まれたナノ粒子のコア内部にさえ、依然として顕著な回転移動性がある。回転の時間目盛りは３０ｎｍのナノ粒子全体が端から端まで回転するには速すぎ、エネルギー転移により偏光解消（depolarization）するには遅すぎる。さらに、曲線の振幅（ｒ（０））が二光子異方性の理論的振幅（ｒ（０）＝０．５７）に近く、これはエネルギー転移に通常関連する高速偏光解消がないことを示唆している。
従って異方性の減衰は、シリカ媒体内での色素分子の回転障害によるものであると思われる。各構造について計量した異方性を図４Ｃに示す。密集コア−殻ナノ粒子が最も速く、回転障害が最も少なく(図４Ｃ）、次が膨張コア−殻粒子であり（図４Ｃ）、最後が均質ナノ粒子である（図４Ｃ）。この回転時間は非放射速度定数への単調な逆の依存があり（表ＩＩＩ）、シリカナノ粒子における限定的な移動性が非放射減衰の抑制の原因であることを示唆している。
ナノ粒子合成用物質
無水エタノール（アルドリッチ社（Aldricｈ））、テトラヒドロフラン（アルドリッチ社）、水酸化アンモニウム（フルカ社（Fluka）、２８％）、テトラエトキシシラン（アルドリッチ社、９８％）、３−アミノプロピル・トリエトキシシラン（アルドリッチ社、９９％）、３−メルカプトプロピル・トリエトキシシラン（ジェレスト社（Gelest）、９９％）、テトラメチルローダミン−５−（及び−６−）−イソチオシアネート＊混合異性体＊（ＴＲＩＴＣ）（モレキュラープローブ社（Molecular Probes）、８８％）、Alexa Fluor（アレクサ・フルーア）（登録商標）４８８ Ｃ５メレイミド（モレキュラープローブ社、９７％）、Alexa Fluor（アレクサ・フルーア）（登録商標）４８８ カルボン酸、及びスクシンイミジル・エステル（モレキュラープローブ社、≧５０％）。
コア（蛍光種）ナノ粒子の調製全般
水、アンモニア、及び溶媒の量は、メスシリンダーで測定した。蛍光種粒子の合成は、１Ｌのエルレンマイヤーフラスコで実施し、磁気テフロン（登録商標）で被覆した撹拌子を用いて、約６００ｒｐｍで撹拌した。脱イオン水とアンモニア溶液をエタノールに添加し、撹拌した。約２ｍＬの反応性色素前駆体を溶解した約４２５マイクロモルのＡＰＴＳを含むエタノール又はＴＨＦを、反応槽に添加した。所望の構造に応じて、得られた混合物を、露光を最小限にするためにアルミホイルで覆った反応槽で室温で１〜１２時間撹拌すると、蛍光種粒子混合物が得られた。テトラヒドラフラン（ＴＨＦ）と無水エタノール（ＥｔＯＨ）を窒素下で蒸留した。有機色素を約−２０℃の保存温度から室温に戻してからグローブボックスに入れた。
コア（蛍光種）粒子上のシリカ殻の調製全般
シリカ殻の被覆及び成長工程は、シリカ形成モノマーであるテトラエトキシシラン（ＴＥＯＳ）を添加しながら、溶液のイオン強度の急激な変化を避けるために、エタノール、メタノールや、イソプロパノールなどの溶媒を規則的に添加して、上で述べた蛍光種粒子反応混合物中で行った。これにより合成中の粒子密集が避けられ、粒径分布を広げることができる。
蛍光ナノ粒子の特性決定
結果的に得られる蛍光ナノ粒子の粒径と粒径分布を、電子顕微鏡検査（ＳＥＭ）と蛍光相関分光法（ＦＣＳ）により特性決定した。
蛍光相関分光法
ＦＣＳ測定は全て、特別注文の二光子励起に基づくＦＣＳ顕微鏡で行った。この装置は、約１００ｆｓのパルス幅、Ａｒ＋イオンレーザー（スペクトラ・フィジックス社（Spectra Physics）、パロアルト、カリフォルニア州）で励起した８０ＭＨｚのｒｅｐ速度を有するＴｉ：サファイア発振器から成る。ビームはバイオラッドＭＲＣ６００共焦点スキャンボックス（ハーキュリーズ、カリフォルニア州）にツァイスのＡｘｉｏｖｅｒｔ３５倒立顕微鏡を接続して位置を決めた。励起光はツァイス６３×Ｃ−アポクロマート水浸レンズ（Ｎ．Ａ．＝１．２）で焦点を合わせ、発光は同じレンズで集光した。蛍光は６７０ＤＣＬＰダイクロイックで励起光から分離し、ＧａＡｓｐ光子計数ＰＭＴまでＨＱ５７５／１５０発光フィルターを通過させた。その結果得られた光子流をＡＬＶ６０１０ 多重タウ自己相関器でデジタル処理で自己相関させた。
自己相関関数G（τ）は、次のとおり定義する。
Figure 2006514708
式中、F（τ）は遅延時間τでのその体積から得られる蛍光であり、山括弧は集合平均を表し、δF（τ）＝F（τ）−＜F（τ）＞である。
フィッティング関数は、一要素三次元拡散についての標準関数である。
Figure 2006514708
式中、ｗｘｙとｗｚはそれぞれ二光子焦点体積の横の長さ、縦の長さであり、Nは焦点体積中の拡散種の数であり、Dは拡散係数である。ＦＣＳ測定は全て、９００ｎｍの励起波長で行った。この波長では、横の長さは０．２４８マイクロメートルであると測定され、縦の長さは０．６４０マイクロメートルであると測定された。濃度較正はローダミングリーンの標準試料から１．４３ｎＭ／粒子（即ち、１．４３ｎＭに対しＮ＝１）と決定された。
光退色
光退色の実験用に、３０μＬの試料を３ｍｍ光路長クォーツキュベット（スタルナ社（Starna））内で連続的に照射した。励起は、出力が５Ｗ、スポットサイズが１．５ｃｍのアルゴンレーザーからの（５１４ｎｍの）レーザー線であった。試料は５１４ｎｍの励起波長で類似の吸光度を持つように調製した。
蛍光は、ＨＱ６０５／９０発光フィルターを通して励起に対して直角に集光し、バイアルカリＰＭＴ（ＨＣ１２５−０２、ハママツ）で記録した。
以下の実施例は説明の目的でのみ提供するものであり、本発明を限定するものではない。
Figure 2006514708
２１℃で計算した流体半径を表Ｉに挙げる。各ナノ粒子曲線は単一の拡散係数に適合し、流体サイズは測定の誤差範囲内で等しい（ｒ＝１５、１７ｎｍ）。コア中間体については、膨張コア（ｒ＝２．９ｎｍ）が密集コア（ｒ＝２．２ｎｍ）よりも僅かに大きい。
Figure 2006514708
自己相関の振幅により拡散種の数が得られ、平均計数率は光学的に定めた焦点体積から回収した光子の尺度である。このデータから、各拡散種に対する分子当たりの計数率が得られ、これがプローブの輝度の直接的な尺度となる。
遊離色素を超える粒子の全体的な輝度の上昇は、表ＩＩの輝度因子に示すように、ナノ粒子内部のＴＲＩＴＣ等価物の数と色素の相対的量子効率向上の積である。しかしながら、ＴＲＩＴＣ等価物の測定時の不正確さを考慮すると、比較用に、ＦＣＳから測定した粒子あたりの計数（表ＩＩ、計数／粒子）など、追加の独立した輝度の尺度を持つことが必要である。輝度因子はＦＣＳからの独立した輝度測定に類似し（表ＩＩ、最後２縦列を比較）、異なる構造にわたる輝度の蛍光は両方の方法で同じである。輝度因子は、恐らくは色素等価物の測定時の誤差のため、ＦＣＳで測定した値を一貫して過大評価する。
量子効率の向上は、一般的に、放射速度（Ｋｒ）の増加、非放射速度（Ｋｎｒ）の減少、又はその両方による可能性がある。これらの因子の相対的寄与率は、次式により、蛍光寿命（τｆ）と量子効率（φ）に関連する。
Figure 2006514708
Figure 2006514708
全ての出版物、特許、及び特許文献は、参照により個別にインコーポレートするかのように、参照により本明細書にインコーポレートされる。本発明は、様々な特異的及び好ましい実施態様及び技術を参照して説明されてきた。しかしながら、本発明の精神と範囲にありながら、多くの変形及び修正が行えることは理解されるべきである。
図１は、異なるシリカナノ粒子構造の概略を表す図である。図１Ａは密集コア−殻ナノ粒子、図１Ｂは膨張コア−殻ナノ粒子、図１Ｃは均質ナノ粒子を表す。密集コア−殻構造及び膨張コア−殻構造のシリカ殻は、色素分子が一つもないシリカを含む。均質ナノ粒子は、マトリックス中にシリカと色素の複合体を含む。 図２Ａは、ナノ粒子及び合成中間体の蛍光相関分光法を表す図である。 図２Ｂは、ナノ粒子及び合成中間体の蛍光相関分光法を表す図である。 図２Ｃは、ナノ粒子及び合成中間体の蛍光相関分光法を表す図である。 図２Ｄは、９００ｎｍ、１．２ｍＷの励起値に基づく密集コア−殻構造の合成段階の輝度（計数／粒子／秒）値を表す図である。 図２Ｄは、９００ｎｍ、１．２ｍＷの励起値に基づく膨張コア−殻構造の合成段階の輝度（計数／粒子／秒）値を表す図である。 図２Ｄは、９００ｎｍ、１．２ｍＷの励起値に基づく均質構造の合成段階の輝度（計数／粒子／秒）値を表す図である。 図３Ａはコア−殻構造によって達成される量子効率向上を表す定常状態分光法を示す。図３Ａは、密集コア−殻ナノ粒子の吸光と蛍光を表す。吸光値は光学密度の単位である。蛍光は使用した有機色素、ＴＲＩＴＣと比較して目盛を定めた。 図３Ｂはコア−殻構造によって達成される量子効率向上を表す定常状態分光法を示す。図３Ｂは、膨張コア−殻ナノ粒子の吸光と蛍光を表す。吸光値は光学密度の単位である。蛍光は使用した有機色素、ＴＲＩＴＣと比較して目盛を定めた。 図３Ｃはコア−殻構造によって達成される量子効率向上を表す定常状態分光法を示す。図３Ｃは、均質ナノ粒子の吸光と蛍光を表す。吸光値は光学密度の単位である。蛍光は使用した有機色素、ＴＲＩＴＣと比較して目盛りを定めた。 図３Ｄはコア−殻構造によって達成される量子効率工場を表す定常状態分光法を示す。図３Ｄは、密集コア−殻ナノ粒子の吸光と励起の比較を表す。 図３Ｅはコア−殻構造によって達成される量子効率増進を表す定常状態分光法を示す。図３Ｅは、膨張コア−殻ナノ粒子の吸光と励起の比較を表す。 図３Ｆはコア−殻構造によって達成される量子効率増進を表す定常状態分光法を示す。図３Ｆは、均質ナノ粒子の吸光と励起の比較を表す。 図４Ａは、ナノ粒子の時間分解蛍光を表す図である。図４Ａは、均質ナノ粒子、膨張コア−殻ナノ粒子、密集コア−殻ナノ粒子、及びＴＲＩＴＣ色素の正規化蛍光減衰を表す。 図４Ｂは、ナノ粒子の時間分解蛍光を表す図である。図４Ｂは、膨張コア−殻ナノ粒子、密集コア−殻ナノ粒子、均質ナノ粒子、及びＴＲＩＴＣ色素の蛍光異方性を表す。 図４Ｃは、ナノ粒子の時間分解蛍光を表す図である。図４Ｃは、図４Ｂに示すナノ粒子曲線の正規化蛍光異方性を表す。蛍光寿命（τｆ）と回転寿命（θ）の平均フィット値を表ＩＩＩにまとめた。

Claims (13)

  1. 有機コア及びシリカ殻を含む蛍光単分散ナノ粒子であって、コア−殻の化学的性質、組成、及び構造が遊離色素及びコアと比べて光物理的特性の向上をもたらすナノ粒子。
  2. 光物理学的特性の向上が、輝度、蛍光量子効率、光安定性、又はそれらの組合せの向上を含むものである、請求項１に記載の蛍光単分散ナノ粒子。
  3. コアが、シリカ殻に囲まれた密集コアを含むものである、請求項１に記載の蛍光単分散ナノ粒子。
  4. コアが、シリカ殻に囲まれた膨張コアを含むものである、請求項１に記載の蛍光単分散ナノ粒子。
  5. コアが、シリカ殻に囲まれた中で低密度に包埋された色素を有する均質な粒子を含むものである、請求項１に記載の蛍光単分散ナノ粒子。
  6. 均質粒子がシリカ殻に囲まれていないものである、請求項５に記載の蛍光単分散ナノ粒子。
  7. 密集コアの半径が２．５ｎｍよりも小さいものである、請求項１に記載の蛍光ナノ粒子。
  8. 膨張コアの半径が２．５ｎｍよりも大きいものである、請求項１に記載の蛍光ナノ粒子。
  9. コア−殻の化学的性質、組成、及び構造がナノ粒子の光安定特性の制御能力を提供するものである、請求項１に記載の蛍光ナノ粒子。
  10. 密集コア構造を有する蛍光単分散ナノ粒子を製造する方法であって、
    蛍光化合物とオルガノシラン化合物を混合して色素前駆体を形成する工程、
    その結果得られた色素前駆体を水溶液と混合して密集蛍光コアを形成する工程、及び、
    その結果得られた密集コアをシリカ前駆体と混合して密集コア上にシリカ殻を形成し、蛍光単分散ナノ粒子を得る工程
    を含む方法。
  11. 膨張コア構造を有する蛍光単分散ナノ粒子を製造する方法であって、
    蛍光化合物とオルガノシラン化合物を混合して色素前駆体を形成する工程、
    その結果得られた色素前駆体をシリカ前駆体と共縮合して膨張蛍光コアを形成する工程、及び、
    その結果得られた膨張コアをシリカ前駆体と混合して膨張コア上にシリカ殻を形成し、蛍光単分散ナノ粒子を得る工程
    を含む方法。
  12. 蛍光単分散均質ナノ粒子を製造する方法であって、
    蛍光化合物とオルガノシラン化合物を混合して色素前駆体を形成する工程、及び、
    その結果得られた色素前駆体をシリカと共縮合して均質な蛍光単分散ナノ粒子を形成する工程
    を含む方法。
  13. 請求項１２に記載の方法であって、結果的に得られた均質ナノ粒子をシリカと共縮合して均質ナノ粒子上にシリカ殻を形成し、蛍光単分散ナノ粒子を得る方法。
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