WO2003060037A1 - Silica sphere containing fluorescent dye molecule - Google Patents
Silica sphere containing fluorescent dye molecule Download PDFInfo
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- WO2003060037A1 WO2003060037A1 PCT/JP2001/011572 JP0111572W WO03060037A1 WO 2003060037 A1 WO2003060037 A1 WO 2003060037A1 JP 0111572 W JP0111572 W JP 0111572W WO 03060037 A1 WO03060037 A1 WO 03060037A1
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
- C09B67/00—Influencing the physical, e.g. the dyeing or printing properties of dyestuffs without chemical reactions, e.g. by treating with solvents grinding or grinding assistants, coating of pigments or dyes; Process features in the making of dyestuff preparations; Dyestuff preparations of a special physical nature, e.g. tablets, films
- C09B67/0001—Post-treatment of organic pigments or dyes
- C09B67/0004—Coated particulate pigments or dyes
- C09B67/0008—Coated particulate pigments or dyes with organic coatings
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
- C09B67/00—Influencing the physical, e.g. the dyeing or printing properties of dyestuffs without chemical reactions, e.g. by treating with solvents grinding or grinding assistants, coating of pigments or dyes; Process features in the making of dyestuff preparations; Dyestuff preparations of a special physical nature, e.g. tablets, films
- C09B67/0033—Blends of pigments; Mixtured crystals; Solid solutions
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1003—Carbocyclic compounds
- C09K2211/1014—Carbocyclic compounds bridged by heteroatoms, e.g. N, P, Si or B
Definitions
- the present invention relates to a fluorescent dye molecule-containing silica sphere useful as a detection system reagent.
- Conventional technology :
- silica spheres containing fluorescent dye molecules therein have been proposed.
- FITC fluorescein isothiocyanate
- APS 3-aminopropyl orthosilicate
- quenching occurs inside the silica sphere at the highest concentration.
- the present invention has been made in view of the above-mentioned problems of the related art, and has an object to provide a fluorescent dye molecule-containing siliceous sphere having excellent manufacturing stability and excellent performance.
- the present inventors have conducted intensive studies in order to achieve the above object, and as a result, have found that it is extremely effective to introduce a specific compound into silica spheres containing fluorescent dye molecules, and have completed the present invention.
- the present invention is a fluorescent dye molecule-containing siliceous sphere prepared using a fluorescent dye molecule represented by the following structural formula (1).
- R is a substance that can have NH S in the side chain.
- R is a substance selected from FITC (fluorescein isothiocyanate), dyes, magnetic substances and free radicals.
- FITC fluorescein isothiocyanate
- FI TC—CO—NH— (CH 2 ) 3 —S i— (C 2 H 5 ⁇ ) 3 ( ⁇ ) is preferred.
- FITC represents fluorescein isothiocynate.
- R is a substance that can have NHS in the side chain, such as other dyes such as rhodamine, magnetic substances such as iron oxide, and stable free radicals such as spin labeling agents. All included.
- the present invention relates to the process of producing FITC-APS-silica, a silica sphere containing fluorescent dye molecules, which has been conventionally proposed.
- FITC—NHS with NHS (N-hydroxysuccinimide ester) as the chain
- FITC—CO— is used for its strong binding ability between the amine group of APS and the CO group of FITC-NH.S. This was prepared from APS, which significantly improved its production stability and the like.
- the method for producing the fluorescent dye molecule-containing siliceous sphere of the present invention may be the same as that of the above-mentioned prior art. That is, a fluorosynsothiothionate or a biotin molecule having an N-hydroxysuccinimide ester in a side chain is used as a raw material, and a silane coupling agent, 3-aminopropyl orthosilicate, is used.
- Fluorescent dye molecule-containing siliceous spheres can be manufactured using tetraorthosilicate.
- a DMSO solution of FITC-CO-APS is added to 6 ml of water, and the ratio of 20 ml of ethanol to an aqueous solution is set to 4: 1.
- the size of the obtained silica sphere can be freely adjusted from several nm in diameter to several hundred nm, and even to the order of m. .
- the obtained silica spheres are purified by removing coexisting ions using a well-known ultrafiltration method, or adjusted to a desired particle size distribution.
- the fluorescent dye molecule-containing silica spheres of the present invention can have a size of several nanometers and several tens of nanometers.
- NHS a detection system reagent that does not easily come off when bound to proteins and is not easily quenched.
- the conventional “FI TC_NHS” molecule has poor stability, so the number of FI TC that binds to the antibody protein is about 4 molecules, the sensitivity is low, and it can be applied only to a fixed form on a petri dish.
- the silica spheres containing the fluorescent dye molecules of the present invention have a maximum of several thousand FITCs in the silica spheres without concentration quenching. It has an excellent effect of strong light emission and high sensitivity.
- silica is chemically inert and easily modified, so that, for example, the surface can be made mesoporous or a specific protein can be newly added. It is possible to immobilize the receptor molecule for binding to the surface.
- FIG. 1 is a view showing an absorption spectrum of a fluorescent dye molecule-containing siliceous sphere (A: 1st Growth, B: 2nd Growth) prepared in Example 1 in an aqueous solution.
- FIG. 2 is a diagram showing a fluorescence decay curve of the fluorescent dye molecule-containing silica spheres (A: 1st Growth, B: 2nd Growth) prepared in Example 1.
- FIG. 3 is a TEM photograph (X 90,000) of the fluorescent pigment molecule-containing silica sphere (1st Growth) prepared in Example 1.
- FIG. 4 is a TEM photograph (X 15,000) of the fluorescent pigment molecule-containing silica spheres (2st Growth) prepared in Example 1.
- a fluorescein isothiocyanate or a biotin molecule having an N-hydroxysuccinimide ester in the side chain is used as a raw material.
- a silane coupling agent 3-aminopropyl orthosilicate
- StSber that is, using tetraorthosilicate in a 4: 1 ethanol aqueous solution at room temperature in the air by hydrolysis and polycondensation of tetraorthosilicate in aqueous ammonia in ethanol.
- silica spheres containing fluorescent dye molecules were produced.
- piotin-silica particles containing a fluorescein isothiocyanate molecule were prepared, and the same observation was carried out. Similar aggregation was observed. The protein label by silica particles was confirmed.
- FIG. 1 shows the absorption spectrum of the silica spheres containing fluorescent dye molecules (A: Ist Growth, B: 2nd Growth) prepared in Example 1 in an aqueous solution.
- the color of the 1st Glo wt h solution was yellow with strong fluorescence, and was stable even after one month. This indicates that the solution pH at the time of sample preparation was strongly alkaline, and was present inside the silica spheres in that state.
- a typical absorption peak was observed at 490 nm, and a scattering effect based on large particles was observed in the absorption spectrum of the second glass, in which the absorption increased toward the short wavelength side.
- the concentration of fluorescein isothiocyanate estimated from the molecular extinction coefficient was 0.075 mM in the case of 1st Growth. Furthermore, when the fluorescence spectra were measured, both had typical fluorescence peaks.
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- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
Description
明 細 蛍光色素分子含有シリカ球 発明の属する技術分野： TECHNICAL FIELD The technical field to which the present invention pertains:
本発明は、 検出系試薬として有用な螢光色素分子含有シリカ球に関する。 従来の技術： The present invention relates to a fluorescent dye molecule-containing silica sphere useful as a detection system reagent. Conventional technology:
近年、 蛍光色素を含む微粒子を利用した生化学的検査手法が各種研究されてい る。 例えば、 パッカード社によりアルファスクリーン技法が商品化されている。 これは、直径 2 5 0 n mのラテックス製ドナーとァクセクタ一ビーズ（登録商標） を使用したもので、 ドナービーズとァクセクタ一ビーズが結合した後、 ドナービ ーズをレーザーで励起すると、 内部の蛍光分子からの蛍光で一重項酸素分子が生 成し、 これがァクセクタ一ビーズ中の蛍光物質と化学反応して化学発光を生じ、 これを観測するというものである。 In recent years, various biochemical inspection methods using fine particles containing a fluorescent dye have been studied. For example, Packard has commercialized the AlphaScreen technique. This method uses a latex donor with a diameter of 250 nm and an X-Axease-Bead (registered trademark). After the donor and the X-Axe-A-Bead are combined, when the donor bead is excited by a laser, internal fluorescent molecules are generated. A singlet oxygen molecule is generated by the fluorescence from this, and this chemically reacts with the fluorescent substance in the vaccinia beads to generate chemiluminescence, which is observed.
一方、 内部に蛍光色素分子を入れたシリカ球の製法も各種提案されており、 代 表的なものとして、 予め A P S ( 3—ァミノプロピルオルソシリゲート） に直接 F I T C (フルォロセインイソチオシァネート） を結合させて調製したものがあ り （J. P ys. Chem. B 1999， 103， 1408-1415) 、 この種の製法の中で最も高濃度 の蛍光色素分子をシリカ球内部に保持している。 しかし、 その最大濃度では、 シ リカ球内部で消光が起こることも報告されている。 このように、 蛍光色素分子を シリカで囲うことで、 外部因子による消光 （生化学的高分子等による励起工ネル ギ一の吸収） がなく、 各種検査に応用されることが期待されている。 On the other hand, various methods for producing silica spheres containing fluorescent dye molecules therein have been proposed. As a typical example, FITC (fluorescein isothiocyanate) is directly added to APS (3-aminopropyl orthosilicate) in advance. (J. Pys. Chem. B 1999, 103, 1408-1415), and the highest concentration of fluorescent dye molecules in this type of process is placed inside silica spheres. keeping. However, it has been reported that quenching occurs inside the silica sphere at the highest concentration. By surrounding the fluorescent dye molecules with silica in this way, there is no quenching due to external factors (absorption of excitation energy by biochemical polymers, etc.), and it is expected to be applied to various tests.
ところが、本発明者らが更に検討を進めた結果、上記のシリ力球の製造上、 FITC . と APSの結合性が高くなく、 製造効率が悪いと共に、 得られた粒子サイズが単 —で数百ナノメーターである等の欠点があった。 発明の開示 However, as a result of further investigations by the present inventors, FITC. In addition, the binding efficiency between APS and APS was not high, the production efficiency was poor, and the obtained particle size was simple and several hundred nanometers. Disclosure of the invention
本発明は、 上記従来技術の課題に鑑みなされたものであり、 製造上の安定性に 優れ、 性能にも優れた蛍光色素分子含有シリ力球の提供を目的とする。 The present invention has been made in view of the above-mentioned problems of the related art, and has an object to provide a fluorescent dye molecule-containing siliceous sphere having excellent manufacturing stability and excellent performance.
本発明者らは、 かかる目的を達成するため鋭意検討した結果、 蛍光色素分子含 有シリカ球に特定の化合物を導入することが極めて有効であることを見出し、 本 発明を完成するに至った。 The present inventors have conducted intensive studies in order to achieve the above object, and as a result, have found that it is extremely effective to introduce a specific compound into silica spheres containing fluorescent dye molecules, and have completed the present invention.
即ち、 本発明は下記構造式 (1) で示される蛍光色素分子を使って調整した蛍光 色素分子含有シリ力球である。 That is, the present invention is a fluorescent dye molecule-containing siliceous sphere prepared using a fluorescent dye molecule represented by the following structural formula (1).
R-CO-NH- (CH2) 3-S i - (C2HsO) 3 (1) R-CO-NH- (CH 2 ) 3 -S i-(C 2 H s O) 3 (1)
式中、 Rは NH Sを側鎖に有することができる物質である。 Wherein R is a substance that can have NH S in the side chain.
好ましくは、 Rは F I TC (フルォロセインイソチオシァネート） 、 色素、 磁 性物質およびフリーラジカルより選ばれた物質である。 Preferably, R is a substance selected from FITC (fluorescein isothiocyanate), dyes, magnetic substances and free radicals.
特に F I TC— CO— NH— （CH2) 3— S i— (C2H5〇） 3 (Γ) が好ましい。 式 (Γ) 中、 F I TCはフルォロセインイソチオシァネートを示す。 In particular, FI TC—CO—NH— (CH 2 ) 3 —S i— (C 2 H 5 〇) 3 (Γ) is preferred. In the formula (Γ), FITC represents fluorescein isothiocynate.
F I TCは 1例であって、 その代わりにローダミンな'どの他の色素、 酸化鉄な どの磁性物質、 スピンラベル剤などの安定フリーラジカルなど、 Rは NHSを側 鎖に有することができる物質をすベて含む。 発明の詳細な説明： FITC is just one example, and instead R is a substance that can have NHS in the side chain, such as other dyes such as rhodamine, magnetic substances such as iron oxide, and stable free radicals such as spin labeling agents. All included. DETAILED DESCRIPTION OF THE INVENTION:
以下に本発明を詳しく説明する。 本発明は、 従来提案されている蛍光色素分子 含有シリカ球である 「F I TC— AP S—シリカ」 の製造過程で、 F I TCに側 鎖として NHS (N—ハイドロキシサクシンイミドエステル） を持つ F I TC— NHSを使用し、 APSのァミン基と F I TC一 NH.Sの CO基との強固な結合 能を利用して F I TC— CO— AP Sを調製したものであり、 これによりその製 造安定性等を顕著に改善したものである。 Hereinafter, the present invention will be described in detail. The present invention relates to the process of producing FITC-APS-silica, a silica sphere containing fluorescent dye molecules, which has been conventionally proposed. Using FITC—NHS with NHS (N-hydroxysuccinimide ester) as the chain, FITC—CO— is used for its strong binding ability between the amine group of APS and the CO group of FITC-NH.S. This was prepared from APS, which significantly improved its production stability and the like.
本発明の蛍光色素分子含有シリ力球の製造方法は、 上記従来技術のものと同様 でよい。 即ち、 N—ハイドロキシサクシンイミドエステルを側鎖に持つフルォロ セィンィソチオシァネ一トまたはピオチン分子を原料として使用し、 これとシラ ンカップリング剤である 3—ァミノプロピルオルソシリケー卜の結合により生成 した化合物を核として、 St erの方法に従って、 即ち、 エタノール水溶液中での テトラオルソシリゲートのアンモニアによる加水分解と重縮合により、 室温で大 気下、 4 ： 1のエタノール水溶液中で、 テトラオルソシリケートを使って蛍光色 素分子含有シリ力球を製造することができる。 The method for producing the fluorescent dye molecule-containing siliceous sphere of the present invention may be the same as that of the above-mentioned prior art. That is, a fluorosynsothiothionate or a biotin molecule having an N-hydroxysuccinimide ester in a side chain is used as a raw material, and a silane coupling agent, 3-aminopropyl orthosilicate, is used. Using the compound formed by the bond as a nucleus, according to the method of Ster, that is, by hydrolysis and polycondensation of tetraorthosilicate in ammonia in an aqueous ethanol solution at room temperature under atmospheric pressure, in a 4: 1 aqueous ethanol solution Fluorescent dye molecule-containing siliceous spheres can be manufactured using tetraorthosilicate.
より詳細には、 F I TC— CO— AP Sの DMSO溶液 5m 1を、 水 6mlに 加えて、 エタノール 20mlと水溶液の比率を 4 ： 1として、 これにテトラオル ソシリゲート 0. 3m 1と約 30%のアンモニア水 2 m 1を加えて、 一日撹拌し ながら放置する。 In more detail, 5 ml of a DMSO solution of FITC-CO-APS is added to 6 ml of water, and the ratio of 20 ml of ethanol to an aqueous solution is set to 4: 1. Add 2 ml of aqueous ammonia, and stir for one day.
この場合、 テトラオルソシリケートの濃度とこの熟成の回数を変えることによ つて、 得られるシリカ球のサイズを、 直径数 nmから数百 nmへ、 更には mォ ーダ一へと自在に調整できる。 In this case, by changing the concentration of tetraorthosilicate and the number of times of ripening, the size of the obtained silica sphere can be freely adjusted from several nm in diameter to several hundred nm, and even to the order of m. .
また、 得られたシリカ球については、 周知のウルトラフィルトレーションメソ ッドを用いて共存イオンを取り除いて精製したり、 希望する粒子径分布に調整す る。 The obtained silica spheres are purified by removing coexisting ions using a well-known ultrafiltration method, or adjusted to a desired particle size distribution.
本発明の蛍光色素分子含有シリカ球は、 上記の通り、 数ナノメーターサイズと 数十ナノメーターサイズとすることができ、 その表面に NHSを調製することで タンパク質に結合した場合に外れにくく、 消光を受けにくい検出系試薬として有 用である。 また、 従来の 「F I TC_NHS」 分子は安定性が悪いため、 抗体の タンパク質に結合する F I TCは 4分子程度であり、 感度が低く、 シャーレ上の 決まった形のものにしか適用できない、 即ち、 実用性能に劣るものであつたのに 対し、 本発明の蛍光色素分子含有シリカ球は、 シリカ球内に最大で数千個の F I T Cが濃度消光することなく密集しているため、 レーザー照射したときの発光が 強く、 感度が高いという優れた効果を有する。 As described above, the fluorescent dye molecule-containing silica spheres of the present invention can have a size of several nanometers and several tens of nanometers. By preparing NHS on the surface, This is useful as a detection system reagent that does not easily come off when bound to proteins and is not easily quenched. In addition, the conventional “FI TC_NHS” molecule has poor stability, so the number of FI TC that binds to the antibody protein is about 4 molecules, the sensitivity is low, and it can be applied only to a fixed form on a petri dish. In contrast to practical performance, the silica spheres containing the fluorescent dye molecules of the present invention have a maximum of several thousand FITCs in the silica spheres without concentration quenching. It has an excellent effect of strong light emission and high sensitivity.
また、 本発明の蛍光色素分子含有シリカ球の場合、 シリカは、 化学的に不活性 であると共に、 その修飾が容易であることから、 例えば、 その表面をメソポーラ スにしたり、 新たに特定のタンパク質と結合するためのァクセプター分子を表面 に固定することが可能である。 図面の簡単な説明： In the case of the silica sphere containing the fluorescent dye molecule of the present invention, silica is chemically inert and easily modified, so that, for example, the surface can be made mesoporous or a specific protein can be newly added. It is possible to immobilize the receptor molecule for binding to the surface. BRIEF DESCRIPTION OF THE DRAWINGS:
図 1は、 実施例 1で調製した蛍光色素分子含有シリ力球 （A： 1 st Gr ow t h, B： 2nd G r ow t h) の水溶液中の吸収スペクトルを示す図である。 図 2は、 実施例 1で調製した蛍光色素分子含有シリカ球 （A： 1st Gr ow t h, B： 2nd Gr owt h) の蛍光減衰曲線を示す図である。 FIG. 1 is a view showing an absorption spectrum of a fluorescent dye molecule-containing siliceous sphere (A: 1st Growth, B: 2nd Growth) prepared in Example 1 in an aqueous solution. FIG. 2 is a diagram showing a fluorescence decay curve of the fluorescent dye molecule-containing silica spheres (A: 1st Growth, B: 2nd Growth) prepared in Example 1.
図 3は、 実施例 1で調製した蛍光色素分子含有シリカ球 （ 1 st Gr owt h) の T EM写真 (X 90,000) である。 FIG. 3 is a TEM photograph (X 90,000) of the fluorescent pigment molecule-containing silica sphere (1st Growth) prepared in Example 1.
図 4は、 実施例 1で調製した蛍光色素分子含有シリカ球 （2st Gr owt h) の T EM写真 （X 15,000)である。 実施例 FIG. 4 is a TEM photograph (X 15,000) of the fluorescent pigment molecule-containing silica spheres (2st Growth) prepared in Example 1. Example
以下、 実施例により本発明を更に具体的に説明するが、 本発明はこれらの実施 例に限定されるものではない。 Hereinafter, the present invention will be described more specifically with reference to Examples. It is not limited to the example.
実施例 1 Example 1
N—ハイドロキシサクシンイミドエステルを側鎖に持つフルォロセインイソチ オシァネートまたはピオチン分子を原料として使用し、 シランカップリング剤で ある 3—ァミノプロピルオルソシリケ一卜の結合により生成した化合物を核とし て、 StSberの方法に従って、 即ち、 エタノール水溶液中でテトラオルソシリケー 卜のアンモニア水による加水分解と重縮合により室温で大気下、 4 ： 1のェタノ —ル水溶液中で、 テトラオルソシリケートを使つて蛍光色素分子含有シリカ球を 製造した。 Using a fluorescein isothiocyanate or a biotin molecule having an N-hydroxysuccinimide ester in the side chain as a raw material, a compound formed by bonding of a silane coupling agent, 3-aminopropyl orthosilicate, is used as a core. According to the method of StSber, that is, using tetraorthosilicate in a 4: 1 ethanol aqueous solution at room temperature in the air by hydrolysis and polycondensation of tetraorthosilicate in aqueous ammonia in ethanol. Thus, silica spheres containing fluorescent dye molecules were produced.
このようにして調製した、 ピオチン/シリカ粒子を、 アビジン鎖を持つタンパ ク質と混合したところ、 同試料のピオチンによって、 アビジン一タンパク質の凝 集が観測できた。 When the thus prepared biotin / silica particles were mixed with a protein having an avidin chain, aggregation of avidin-protein could be observed with the same sample of biotin.
また、 同様にして、 フルォロセィンィソチオシァネート分子を含むピオチン一 シリカ粒子を調製し、 同様の観察を行ったところ、 同様の凝集が観測でき、 フル ォロセインイソチオシァネート一シリカ粒子によるタンパク質ラベルが確認でき た。 In the same manner, piotin-silica particles containing a fluorescein isothiocyanate molecule were prepared, and the same observation was carried out. Similar aggregation was observed. The protein label by silica particles was confirmed.
実施例 2 Example 2
実施例 1で調製した蛍光色素分子含有シリカ球 （A : IstGr owt h, B： 2nd Gr owt h) の水溶液中の吸収スペクトルを図 1に示す。 1st G r o wt h溶液の色は強い蛍光を有する黄色を呈しており、 1ヶ月を経ても安定であ つた。 これは、 試料調製時の溶液 pHが強アルカリ性であり、 その状態のまま、 シリカ球の内部に存在していることを示している。 両者とも典型的な吸収ピーク が 490 nmに観測でき、 2nd Gr owt hの吸収スぺクトルは短波長側に向 かって吸収が増大する、 大きな粒子に基づく散乱効果が観測された。 分子吸光係数から推定されるフルォロセインイソチオシァネートの濃度は、 1 st Gr owt hの場合、 0. 075mMであった。 さらに、 蛍光スペクトルを 測定すると、 両者とも典型的な蛍光ピークを有していた。 FIG. 1 shows the absorption spectrum of the silica spheres containing fluorescent dye molecules (A: Ist Growth, B: 2nd Growth) prepared in Example 1 in an aqueous solution. The color of the 1st Glo wt h solution was yellow with strong fluorescence, and was stable even after one month. This indicates that the solution pH at the time of sample preparation was strongly alkaline, and was present inside the silica spheres in that state. In both cases, a typical absorption peak was observed at 490 nm, and a scattering effect based on large particles was observed in the absorption spectrum of the second glass, in which the absorption increased toward the short wavelength side. The concentration of fluorescein isothiocyanate estimated from the molecular extinction coefficient was 0.075 mM in the case of 1st Growth. Furthermore, when the fluorescence spectra were measured, both had typical fluorescence peaks.
また、 蛍光寿命測定の結果 （図 2) によると、 水溶液中でのフリーな状態なも の （3. 8 n s e c) とほぼ同じであったことから、 本試料ではシリカ球内への 濃縮による自己消光は起こっていないことがわかつた。 Also, according to the result of fluorescence lifetime measurement (Fig. 2), it was almost the same as the free state in aqueous solution (3.8 nsec). It turned out that no quenching had occurred.
また、 同試料を透過型電子顕微鏡で観察すると、 数 nmから数百 nmの粒子が 観測でき、 1st Gr owt hの方 （図 3) は表面に凹凸があり、 メソポーラス 様を呈しており、 2nd Gr owt hの方 （図 4) は表面が滑らかであった。 When the same sample was observed with a transmission electron microscope, particles of several nm to several hundreds of nm could be observed. The surface of the 1st glass (Fig. 3) had irregularities and a mesoporous appearance. The surface of the glass (Fig. 4) was smooth.
Claims
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| PCT/JP2001/011572 WO2003060037A1 (en) | 2001-12-27 | 2001-12-27 | Silica sphere containing fluorescent dye molecule |
| JP2003546639A JPWO2003060037A1 (en) | 2001-12-27 | 2001-12-27 | Silica spheres containing fluorescent dye molecules |
| AU2002216422A AU2002216422A1 (en) | 2001-12-27 | 2001-12-27 | Silica sphere containing fluorescent dye molecule |
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Cited By (9)
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| JP2006514708A (en) * | 2002-11-26 | 2006-05-11 | コーネル リサーチ ファウンデーション，インコーポレーティッド | Silica-based fluorescent nanoparticles |
| JP2006300930A (en) * | 2005-03-22 | 2006-11-02 | Univ Of Tokushima | Fluorescent material and X-ray sensitized substrate |
| EP2062946A1 (en) * | 2007-08-13 | 2009-05-27 | Procter & Gamble International Operations SA. | Dye-loaded particles |
| WO2009072657A1 (en) * | 2007-12-06 | 2009-06-11 | The University Of Tokushima | Nanofunctional silica particles and manufacturing method thereof |
| US8168095B2 (en) | 2007-08-13 | 2012-05-01 | The Procter & Gamble Company | Spray-drying process for the manufacture of dye-loaded particles |
| JP2012532211A (en) * | 2009-06-30 | 2012-12-13 | スリーエム イノベイティブ プロパティズ カンパニー | Transparent fluorescent structure with improved fluorescence using nanoparticles, method for producing the same, and use thereof |
| CN103904804A (en) * | 2012-12-28 | 2014-07-02 | 上海微电子装备有限公司 | Coil unit for machine winding and planar motor using the coil unit |
| CN106118636A (en) * | 2016-06-27 | 2016-11-16 | 高大元 | A kind of preparation method of fluorescence silicon oxide nano-particle |
| CN110964341A (en) * | 2019-11-29 | 2020-04-07 | 安徽捷融数据科技有限公司 | Preparation method of natural dye with high washing fastness for cotton products |
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| US4918200A (en) * | 1984-07-16 | 1990-04-17 | Huls America Inc. | Chromogenic and fluorogenic silanes and using the same |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2014133893A (en) * | 2002-11-26 | 2014-07-24 | Cornell Research Foundation Inc | Fluorescent silica-based nanoparticle |
| JP2006514708A (en) * | 2002-11-26 | 2006-05-11 | コーネル リサーチ ファウンデーション，インコーポレーティッド | Silica-based fluorescent nanoparticles |
| JP2010070768A (en) * | 2002-11-26 | 2010-04-02 | Cornell Research Foundation Inc | Fluorescent silica-based nanoparticle |
| JP2011052228A (en) * | 2002-11-26 | 2011-03-17 | Cornell Research Foundation Inc | Fluorescent silica-based nanoparticle |
| JP2006300930A (en) * | 2005-03-22 | 2006-11-02 | Univ Of Tokushima | Fluorescent material and X-ray sensitized substrate |
| EP2062946A1 (en) * | 2007-08-13 | 2009-05-27 | Procter & Gamble International Operations SA. | Dye-loaded particles |
| WO2009022307A3 (en) * | 2007-08-13 | 2009-12-17 | Procter & Gamble International Operations Sa | Dye-loaded particles |
| US8168095B2 (en) | 2007-08-13 | 2012-05-01 | The Procter & Gamble Company | Spray-drying process for the manufacture of dye-loaded particles |
| WO2009072657A1 (en) * | 2007-12-06 | 2009-06-11 | The University Of Tokushima | Nanofunctional silica particles and manufacturing method thereof |
| JP2012532211A (en) * | 2009-06-30 | 2012-12-13 | スリーエム イノベイティブ プロパティズ カンパニー | Transparent fluorescent structure with improved fluorescence using nanoparticles, method for producing the same, and use thereof |
| CN103904804A (en) * | 2012-12-28 | 2014-07-02 | 上海微电子装备有限公司 | Coil unit for machine winding and planar motor using the coil unit |
| CN106118636A (en) * | 2016-06-27 | 2016-11-16 | 高大元 | A kind of preparation method of fluorescence silicon oxide nano-particle |
| CN110964341A (en) * | 2019-11-29 | 2020-04-07 | 安徽捷融数据科技有限公司 | Preparation method of natural dye with high washing fastness for cotton products |
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| JPWO2003060037A1 (en) | 2005-05-19 |
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