TWI465709B - Apparatus for fluorescence enhancement - Google Patents
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Description
本發明係關於螢光增強裝置，特別是關於利用光子晶體之螢光增強裝置。This invention relates to fluorescent enhancement devices, and more particularly to fluorescent enhancement devices utilizing photonic crystals.
螢光檢測技術為環境、生醫、食品...等等常見之檢測應用，然螢光檢測常遭遇螢光強度低而影響螢光檢測效果之困擾。光子晶體具有特殊光能隙，可應用於提高激發光強度、螢光萃取效率，為近年來提高螢光檢測靈敏度方法之一。但目前設計之光子晶體螢光檢測器僅能單次使用，且反應時間長，因此並無法進行多次或連續監測。Fluorescent detection technology is a common detection application for environmental, biomedical, food, etc. Fluorescence detection often suffers from low fluorescence intensity and affects the detection effect of fluorescent detection. The photonic crystal has a special optical energy gap and can be applied to improve the excitation light intensity and the fluorescence extraction efficiency, and is one of the methods for improving the fluorescence detection sensitivity in recent years. However, the photonic crystal fluorescent detector currently designed can only be used in a single use, and the reaction time is long, so it is impossible to perform multiple or continuous monitoring.
目前已知的技術或文獻係將螢光材料直接與光子晶體接觸，藉由該光子晶體之光能隙與螢光材料之螢光波長重疊(部分同時含有與激發光波長重疊之光能隙)，達到螢光強度增強的效果。Cui等人(Anal.Methods,2010,2,448-450.)利用可檢測汞離子之螢光材料與光子晶體混合，該光子晶體之光能隙與螢光波長重疊，達到提高螢光檢測靈敏度。Li等人(Journal of Colloid and Interface Science 356,2011,63-68.)利用兩層不同光能隙之光子晶體，其中一層光能隙與激發光重疊，另一層光能隙與螢光重疊，當螢光材料滲入該光子晶體複合層中，達到提高螢光強度效果。然而上述先前技術的光子晶體感測器皆不具重複使用功能。螢光材料直接與光子晶體接觸造成待測物在光子晶體間質傳的問題，限制檢測與清洗效率，並在反覆使用中破壞光子晶體結構與螢光材料特性，致使光子晶體感 測器無法多次重複使用，亦無連續監測應用能力。另外螢光材料與光子晶體混合，部份發光會被光子晶體反射至光學檢測器相反方向，而降低可被檢測的螢光強度。The currently known technology or literature directly contacts the fluorescent material with the photonic crystal, whereby the optical energy gap of the photonic crystal overlaps with the fluorescent wavelength of the fluorescent material (partially containing a light energy gap overlapping the wavelength of the excitation light) , to achieve the effect of enhanced fluorescence intensity. Cui et al. (Anal. Methods, 2010, 2, 448-450.) use a fluorescent material that can detect mercury ions to mix with a photonic crystal. The photonic energy gap of the photonic crystal overlaps with the wavelength of the fluorescent light to improve the fluorescence detection sensitivity. Li et al. (Journal of Colloid and Interface Science 356, 2011, 63-68.) utilize two photonic crystals of different optical energy gaps, one of which overlaps with the excitation light and the other of which overlaps with the fluorescent light. When the fluorescent material penetrates into the photonic crystal composite layer, the effect of improving the fluorescence intensity is achieved. However, none of the prior art photonic crystal sensors described above have a reusable function. The direct contact of the fluorescent material with the photonic crystal causes the problem of mass transfer of the analyte between the photonic crystals, limits the detection and cleaning efficiency, and destroys the photonic crystal structure and the characteristics of the fluorescent material in the repeated use, resulting in a photonic crystal sensation. The detector cannot be reused multiple times and there is no continuous monitoring application capability. In addition, the phosphor material is mixed with the photonic crystal, and some of the light is reflected by the photonic crystal to the opposite direction of the optical detector, thereby reducing the intensity of the fluorescent light that can be detected.
綜合上述，開發一種可重複使用且可增強螢光強度的光子晶體感測器是目前需努力的目標。In summary, the development of a reusable photonic crystal sensor that enhances fluorescence intensity is currently a goal.
本發明的目的之一係開發一種螢光增強裝置，其可重複使用光子晶體進行感測並可增強螢光強度。One of the objects of the present invention is to develop a fluorescence enhancement device that can re-use a photonic crystal for sensing and enhance fluorescence intensity.
依據本發明之一實施例，一種螢光增強裝置包含光源、螢光材料層、第一光子晶體層及第二光子晶體層。光源可發射激發光，激發螢光材料層中的螢光材料進而發射螢光。第一光子晶體層與第二光子晶體層係設置於螢光材料層之激發光入射方向之下游。第一光子晶體層之第一光子晶體之光能隙與激發光之波長重疊，藉以使得第一光子晶體層反射激發光至螢光材料層。第二光子晶體層之第二光子晶體光能隙與螢光材料之螢光波長重疊，藉以反射螢光材料所發射之螢光。According to an embodiment of the invention, a fluorescent enhancement device includes a light source, a phosphor layer, a first photonic crystal layer, and a second photonic crystal layer. The light source can emit excitation light to excite the fluorescent material in the layer of phosphor material to emit fluorescence. The first photonic crystal layer and the second photonic crystal layer are disposed downstream of the incident direction of the excitation light of the phosphor layer. The optical energy gap of the first photonic crystal of the first photonic crystal layer overlaps with the wavelength of the excitation light, whereby the first photonic crystal layer reflects the excitation light to the phosphor material layer. The second photonic crystal optical energy gap of the second photonic crystal layer overlaps with the fluorescent wavelength of the fluorescent material to reflect the fluorescent light emitted by the fluorescent material.
以下藉由具體實施例配合所附的圖式詳加說明，當更容易瞭解本發明之目的、技術內容、特點及其所達成之功效。The purpose, technical contents, features, and effects achieved by the present invention will become more apparent from the detailed description of the appended claims.
1‧‧‧光源1‧‧‧Light source
2‧‧‧螢光材料層2‧‧‧Fluorescent material layer
21‧‧‧螢光材料21‧‧‧Fluorescent materials
3‧‧‧第一光子晶體層3‧‧‧First photonic crystal layer
4‧‧‧第二光子晶體層4‧‧‧Second photonic crystal layer
5‧‧‧透明基材5‧‧‧Transparent substrate
6‧‧‧載體6‧‧‧ Carrier
圖1及圖2為示意圖顯示本發明之一實施例之一種螢光增強裝置。1 and 2 are schematic views showing a fluorescent enhancement device according to an embodiment of the present invention.
圖3為光譜吸收圖顯示第一及第二光子晶體之光能隙及激發光與螢光材料的波長範圍。3 is a spectral absorption diagram showing the optical energy gap of the first and second photonic crystals and the wavelength range of the excitation light and the fluorescent material.
圖4為實驗數據顯示本發明之一實施例以光子晶體反射激發光與螢光所得之螢光增強結果。Fig. 4 is a graph showing experimental results showing fluorescence enhancement results obtained by reflecting excitation light and fluorescence in a photonic crystal according to an embodiment of the present invention.
圖5為實驗數據顯示本發明之一實施例之水中銅離子濃度對應螢光強度。Fig. 5 is a graph showing experimental data showing that the concentration of copper ions in water according to an embodiment of the present invention corresponds to the fluorescence intensity.
圖6為實驗數據顯示本發明之一實施例之水中銅離子濃度與螢光淬滅量關係。Fig. 6 is a graph showing experimental data showing the relationship between the concentration of copper ions in water and the amount of fluorescence quenching in an embodiment of the present invention.
請參照圖1及圖2，其為示意圖顯示本發明之一實施例之一種螢光增強裝置，包含光源1、螢光材料層2、第一光子晶體層3及第二光子晶體層4。光源1可發射激發光，激發螢光材料層2中的螢光材料21進而發射螢光(圖2之虛線部分)。Please refer to FIG. 1 and FIG. 2 , which are schematic diagrams showing a fluorescent enhancement device according to an embodiment of the present invention, comprising a light source 1 , a phosphor layer 2 , a first photonic crystal layer 3 and a second photonic crystal layer 4 . The light source 1 can emit excitation light to excite the fluorescent material 21 in the phosphor material layer 2 to emit fluorescence (the dotted line portion of Fig. 2).
第一光子晶體層3與第二光子晶體層4係設置於螢光材料層2之激發光入射方向之下游。第一光子晶體層3之第一光子晶體之光能隙與激發光之波長重疊，藉以使得第一光子晶體層3反射激發光至螢光材料層2，如圖1所示，而使螢光材料層2之螢光材料21再度被激發而發射螢光。The first photonic crystal layer 3 and the second photonic crystal layer 4 are disposed downstream of the incident direction of the excitation light of the phosphor layer 2. The optical energy gap of the first photonic crystal of the first photonic crystal layer 3 overlaps with the wavelength of the excitation light, so that the first photonic crystal layer 3 reflects the excitation light to the phosphor material layer 2, as shown in FIG. The phosphor material 21 of the material layer 2 is again excited to emit fluorescence.
第二光子晶體層4之第二光子晶體光能隙與螢光材料21之螢光波長重疊，藉以反射螢光材料21所發射之螢光，如圖2所示，藉以增加螢光強度。The second photonic crystal optical energy gap of the second photonic crystal layer 4 overlaps with the fluorescent wavelength of the fluorescent material 21, thereby reflecting the fluorescent light emitted by the fluorescent material 21, as shown in FIG. 2, thereby increasing the fluorescence intensity.
此處所指的光子晶體是週期性的介電質分佈結構，本發明之第一光子晶體及第二光子晶體可為一維、二維或三維排列。其中在一較佳實施例中，第一光子晶體及第二光子晶體為一維排列，其作用為全反射 膜，藉以反射激發光或螢光。其中一維排列光子晶體可包括布拉格反射層(distributed Bragg reflector,DBR)。The photonic crystal referred to herein is a periodic dielectric distribution structure, and the first photonic crystal and the second photonic crystal of the present invention may be arranged in one, two or three dimensions. In a preferred embodiment, the first photonic crystal and the second photonic crystal are one-dimensionally arranged, and the function is total reflection. Membrane to reflect excitation or fluorescence. The one-dimensional array of photonic crystals may include a distributed Bragg reflector (DBR).
在一實施例中，第一及第二光子晶體係由一微米球自組裝所形成，其中微米球之組成為一有機高分子、一無機成份或其組合。此處的有機高分子可包含但不限於聚苯乙烯系列、聚甲基丙烯酸甲酯系列、聚馬來酸系列、聚乳酸系列、聚胺基酸系列的高分子或其組合。無機成份可包含但不限於矽、鈦、鋯、金、銀、鐵、鋁、銅、鎳金屬、其金屬氧化物或其組合。此外微米球組成可由有機-無機成分所組成，其可包含但不限於碳-矽、碳-鈦、碳-鋯、碳-鋁系列之材質及其組合。In one embodiment, the first and second photonic crystal systems are formed by self-assembly of a micron sphere, wherein the composition of the microspheres is an organic polymer, an inorganic component, or a combination thereof. The organic polymer herein may include, but is not limited to, a polystyrene series, a polymethyl methacrylate series, a polymaleic acid series, a polylactic acid series, a polyamino acid series polymer, or a combination thereof. Inorganic components can include, but are not limited to, tantalum, titanium, zirconium, gold, silver, iron, aluminum, copper, nickel metal, metal oxides thereof, or combinations thereof. In addition, the microsphere composition may be composed of an organic-inorganic component, which may include, but is not limited to, carbon-germanium, carbon-titanium, carbon-zirconium, carbon-aluminum series materials, and combinations thereof.
如圖1所示，第一光子晶體層3係設置於第二光子晶體層4之激發光入射方向之上游。然而第一光子晶體層3與第二光子晶體層4之相對位置可作調整，在本發明之另一實施例中，第二光子晶體層4係設置於第一光子晶體層3之激發光入射方向之上游。As shown in FIG. 1, the first photonic crystal layer 3 is disposed upstream of the incident direction of the excitation light of the second photonic crystal layer 4. However, the relative position of the first photonic crystal layer 3 and the second photonic crystal layer 4 can be adjusted. In another embodiment of the present invention, the second photonic crystal layer 4 is disposed at the excitation of the first photonic crystal layer 3. Upstream of direction.
圖1所示之光源1係設置於第一光子晶體層3與第二光子晶體層4之上方；然而，光源1亦可設置於第一光子晶體層3與第二光子晶體層4之下方或是其他相對位置，只要光源1可射入螢光材料層2及第一光子晶體層3即可。The light source 1 shown in FIG. 1 is disposed above the first photonic crystal layer 3 and the second photonic crystal layer 4; however, the light source 1 may also be disposed under the first photonic crystal layer 3 and the second photonic crystal layer 4 or It is another relative position as long as the light source 1 can be incident on the phosphor layer 2 and the first photonic crystal layer 3.
此外，如圖1所示，本發明之螢光增強裝置包含設置於第二反射層及第一反射層之間的一透明基材5，由此可知第一光子晶體層3與第二光子晶體層4為可分離的。此外，第二光子晶體層4可設置於一載體6之上。In addition, as shown in FIG. 1, the fluorescent enhancement device of the present invention comprises a transparent substrate 5 disposed between the second reflective layer and the first reflective layer, whereby the first photonic crystal layer 3 and the second photonic crystal are known. Layer 4 is separable. Furthermore, the second photonic crystal layer 4 can be disposed on a carrier 6.
在一實施例中，可由兩塊蓋玻片建構一小區域即可界定螢 光材料層2，使其可置放螢光材料21，然而其他界定螢光材料層2之方式亦為可行，只要達成承載螢光材料及透光即可。In one embodiment, a small area can be constructed from two coverslips to define the firefly. The light material layer 2 is such that the fluorescent material 21 can be placed. However, other ways of defining the fluorescent material layer 2 are also feasible, as long as the fluorescent material and the light transmission are achieved.
本項技術領域人士可知螢光材料的多樣性，其可約略分類為有機螢光分子、無機螢光分子，在此未詳列，其中有機螢光分子包括但不限於螢光蛋白(tluorescent proteins)等。A person skilled in the art may know the diversity of fluorescent materials, which may be roughly classified into organic fluorescent molecules and inorganic fluorescent molecules, which are not detailed herein, wherein organic fluorescent molecules include, but are not limited to, tluorescent proteins. .
其中在一實施例中，螢光材料包含一量子點，舉例但不限於CdS、CdSe、CdTe、CdPo、ZnS、ZnSe、ZnTe、ZnPo、MgS、MgSe、MgTe、PbSe、PbS、PbTe、HgS、HgSe或HgTe。In one embodiment, the phosphor material comprises a quantum dot, such as but not limited to CdS, CdSe, CdTe, CdPo, ZnS, ZnSe, ZnTe, ZnPo, MgS, MgSe, MgTe, PbSe, PbS, PbTe, HgS, HgSe. Or HgTe.
第一光子晶體與第二光子晶體之光能隙係藉由微米球之粒徑與有效折射率所調整。The optical energy gap of the first photonic crystal and the second photonic crystal is adjusted by the particle size of the microspheres and the effective refractive index.
請參照上式光能隙公式(T.Endo et al./Sensors and Actuators B 125(2007)589-595)，其由布拉格公式所推導，其中m 指的是衍射等級；λpeak 為衍射峰波長；d111 為(111)平面的間格距離；θ為入射光與折射面法向量之間的夾角；neff 為晶格的平均折射率。其中沿著(111)相緊密填充的面心立方晶體(FCC)結構中，d111 =0.816D，其中D指的是球半徑。Please refer to the above formula for the optical energy gap (T. Endo et al./Sensors and Actuators B 125 (2007) 589-595), which is derived from the Bragg formula, where m refers to the diffraction level; λ peak is the diffraction peak wavelength. d 111 is the inter-frame distance of the (111) plane; θ is the angle between the incident light and the normal of the refractive surface; n eff is the average refractive index of the lattice. Where in the face-centered cubic crystal (FCC) structure closely packed in the (111) phase, d 111 = 0.816D, where D refers to the radius of the sphere.
舉例而言，本發明之螢光增強裝置可用於檢測，應用領域包含但不限於環境、生醫、食品。在一實施例中，可將待測物(未顯示)放置於螢光材料層，以便與螢光材料共反應，進而檢測待測物之濃度。For example, the fluorescent enhancement device of the present invention can be used for detection, and the application field includes, but is not limited to, environment, biomedicine, food. In one embodiment, an object to be tested (not shown) may be placed on the layer of phosphor material to co-react with the phosphor material to detect the concentration of the analyte.
以下通過具體實施例配合附圖詳加說明，可更容易瞭解本發明的目的、技術內容、特點及所達成的功效，並據以實 施，但不能以此限定本發明的保護範圍。The purpose, the technical content, the features and the achieved effects of the present invention can be more easily understood by the following detailed description of the embodiments with reference to the accompanying drawings. This is not intended to limit the scope of the invention.
光子晶體之製備Preparation of photonic crystals
請參照圖3，其為光譜吸收圖顯示第一及第二光子晶體之光能隙及激發光與螢光材料相對應的反射、吸發光波長及強度。第一光子晶體以160nm苯乙烯微米球自組裝之蛋白石，其間隙以二氧化矽(silica)填滿，以下簡稱PC160nm，其用以反射激發光，其中激發光為LED 395 nm。本實施例以重力沉降自組裝法製備三維光子晶體結構，本法所製得之蛋白石(光子晶體)厚度可大於100微米。第二光子晶體以240nm苯乙烯微米球自組裝之蛋白石，其間隙以二氧化矽(silica)填滿，以下簡稱PC240nm其用以反射螢光，其中螢光物質為CdS/ZnS量子點，螢光波峰為554 nm。Please refer to FIG. 3 , which is a spectral absorption diagram showing the optical energy gap of the first and second photonic crystals and the reflection, absorption wavelength and intensity of the excitation light corresponding to the fluorescent material. The first photonic crystal is self-assembled opal with 160 nm styrene microspheres, and the gap is filled with silica, hereinafter referred to as PC160nm, which is used to reflect the excitation light, wherein the excitation light is LED 395 nm. In this embodiment, a three-dimensional photonic crystal structure is prepared by gravity sedimentation self-assembly method, and the thickness of the opal (photonic crystal) prepared by the method can be greater than 100 micrometers. The second photonic crystal is self-assembled opal with 240 nm styrene microspheres, and the gap is filled with silica, hereinafter referred to as PC240nm for reflecting fluorescence, wherein the fluorescent substance is CdS/ZnS quantum dots, fluorescent The peak is 554 nm.
螢光檢測Fluorescence detection
以化學法合成之CdS/ZnS核殼量子點(合成時以巰基乙酸(mercaptoacetic acid)為穩定劑)，作為水中銅離子濃度檢測之螢光感測材料，並以Ocean Optics LED 395nm為激發光源，進行水中銅離子螢光檢測，圖3為LED 395nm激發光與其所激發CdS/ZnS核殼量子點所得螢光光譜圖，圖4為CdS/ZnS核殼量子點以光子晶體反射激發光與螢光所得之螢光增強結果，其中同時加入第一光子晶體PC160及第二光子晶體PC240的組別，相較對照組、僅加入PC160及PC240的組別，明顯增加螢光增強因子效率。Chemically synthesized CdS/ZnS core-shell quantum dots (mercaptoacetic acid as a stabilizer), used as a fluorescent sensing material for copper ion concentration detection in water, and using Ocean Optics LED 395nm as excitation source. The copper ion fluorescence detection in water is carried out. Figure 3 shows the fluorescence spectrum of the 395 nm excitation light and the excited CdS/ZnS core-shell quantum dots. Figure 4 shows the CdS/ZnS core-shell quantum dots reflected by the photonic crystal. The resulting fluorescence enhancement results in which the first photonic crystal PC160 and the second photonic crystal PC240 were simultaneously added, and the efficiency of the fluorescence enhancement factor was significantly increased compared with the control group and only the PC160 and PC240 groups.
圖5為本發明之雙反射光子晶體感測器搭配CdS/ZnS核殼量子點偵測水中銅離子整體結果濃度，圖6為水中銅離子濃度與螢光淬滅 量關係，其中螢光淬滅指的是激發態的螢光分子通過各種外轉換過程失去能量使螢光強度降低的現象。測試結果顯示，本發明可將螢光強度提高23.7倍，且螢光強度隨水中銅離子濃度升高降低，其螢光淬滅量可作為水中銅離子濃度檢測依據。5 is a double-reflection photonic crystal sensor of the present invention combined with a CdS/ZnS core-shell quantum dot to detect the overall concentration of copper ions in water, and FIG. 6 is a copper ion concentration and fluorescence quenching in water. The quantitative relationship, in which fluorescence quenching refers to the phenomenon that the excited fluorescent molecules lose energy by various external conversion processes to reduce the fluorescence intensity. The test results show that the present invention can increase the fluorescence intensity by 23.7 times, and the fluorescence intensity decreases with the increase of the copper ion concentration in the water, and the amount of fluorescence quenching can be used as the basis for detecting the copper ion concentration in the water.
綜合上述，本發明將分別與激發光、螢光波長重疊之複數光子晶體層組合，可建構可置放螢光材料或螢光材料與待測物混合之樣品的小型容器。如此，除可達到增強螢光檢測之螢光強度外，並可使檢測樣品與螢光物質不與光子晶體結構接觸而造成其污染，使得使用光子晶體的檢測器可重複使用多次，並具連續增強螢光效能之檢測能力，藉以擴大光子晶體實務應用之範圍。In summary, the present invention combines a plurality of photonic crystal layers respectively overlapping excitation light and fluorescence wavelength, and constructs a small container capable of placing a sample of a fluorescent material or a fluorescent material mixed with a sample to be tested. In this way, in addition to the enhanced fluorescence intensity of the fluorescent detection, the detection sample and the fluorescent substance can be prevented from being contaminated by contact with the photonic crystal structure, so that the detector using the photonic crystal can be repeatedly used, and has Continuously enhance the detection capability of fluorescent performance to expand the range of practical applications of photonic crystals.
以上所述之實施例僅係為說明本發明之技術思想及特點，其目的在使熟習此項技藝之人士能夠瞭解本發明之內容並據以實施，當不能以之限定本發明之專利範圍，即大凡依本發明所揭示之精神所作之均等變化或修飾，仍應涵蓋在本發明之專利範圍內。The embodiments described above are merely illustrative of the technical spirit and the features of the present invention, and the objects of the present invention can be understood by those skilled in the art, and the scope of the present invention cannot be limited thereto. That is, the equivalent variations or modifications made by the spirit of the present invention should still be included in the scope of the present invention.
1‧‧‧光源1‧‧‧Light source
2‧‧‧螢光材料層2‧‧‧Fluorescent material layer
21‧‧‧螢光材料21‧‧‧Fluorescent materials
3‧‧‧第一光子晶體層3‧‧‧First photonic crystal layer
4‧‧‧第二光子晶體層4‧‧‧Second photonic crystal layer
5‧‧‧透明基材5‧‧‧Transparent substrate
6‧‧‧載體6‧‧‧ Carrier
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| TW201140032A (en) * | 2010-05-11 | 2011-11-16 | Univ Nat Taiwan | Gene inspecting apparatus and gene inspecting method |
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| TW201140032A (en) * | 2010-05-11 | 2011-11-16 | Univ Nat Taiwan | Gene inspecting apparatus and gene inspecting method |
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