I. Gregor, A. Ghosh, Akshita Sharma, Sebastian Isbaner, A. Chizhik, N. Karedla, J. Enderlein
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[3,4] Here, we show that using a graphene layer the localization accuracy of MIET reaches Angstrom accuracy. At such accuracy, minute details such as nanometer-scale roughness of the sample surfaces becomes important.\nFor proof of principle, we determined absolute distances of single molecules from a surface for samples with an a priori well-known sample geometry. We spin-coated fluorescent dye molecules (Atto655) on top of three different substrates with spacer thickness values of 10, 15, and 20 nm, defining the distance of the molecules from the graphene layer. Next, we determined the thickness of supported lipid bilayers (SLBs) by localizing fluorescent dyes attached to lipid head groups in the bottom and top leaflet of the SLB.\nWe have demonstrated that by using graphene as the energy acceptor in MIET, the axial localization accuracy and resolution reaches sub-nanometer levels at photon budgets which are typical in conventional SMLM experiments. An interesting feature of graphene-MIET is that it provides an axial localization accuracy which now surpasses significantly that of lateral localization provided by most SMLM approaches.\n[1] A. I. Chizhik et al. Nat. Photonics 8, 124 (2014).\n[2] S. Isbaner et al. Nano Lett. 18, 2616 (2018).\n[3] N. Karedla et al. ChemPhysChem 15, 705 7 (2014).\n[4] N. Karedla et al. J. Chem. Phys. 148, 204201 (2018).","PeriodicalId":194933,"journal":{"name":"Single Molecule Spectroscopy and Superresolution Imaging XII","volume":"1 1","pages":"0"},"PeriodicalIF":0.0000,"publicationDate":"2019-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Graphene-induced energy transfer: resolving distances at the Angström scale (Conference Presentation)\",\"authors\":\"I. Gregor, A. Ghosh, Akshita Sharma, Sebastian Isbaner, A. Chizhik, N. Karedla, J. Enderlein\",\"doi\":\"10.1117/12.2507939\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"In recent years single-molecule localization super-resolution microscopy (SMLM) has become an indispensable tool for many fields of research. Here, for any image of a single molecule one determines its center position with much higher accuracy than the size of that image itself. A challenge of SMLM is to achieve super-resolution also along the third dimension. Recently, Metal-Induced Energy Transfer or MIET [1,2] was introduced. It exploits the energy transfer from an excited fluorophore to plasmons in a thin metal film. Similar to Forster Resonance Energy Transfer (FRET), this coupling shows a strong distance dependence, but over a range up to 150 nm and enables axial localization of fluorophores with 5-6 nm resolution at a photon budget of 1000 photons. [3,4] Here, we show that using a graphene layer the localization accuracy of MIET reaches Angstrom accuracy. 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引用次数: 0
摘要
近年来,单分子定位超分辨显微镜(SMLM)已成为许多研究领域不可或缺的工具。在这里,对于单个分子的任何图像,人们确定其中心位置的精度要比图像本身的大小高得多。SMLM的一个挑战是在第三维上实现超分辨率。最近引入了金属诱导能量转移(Metal-Induced Energy Transfer,简称MIET)[1,2]。它利用从激发的荧光团到金属薄膜中的等离子体的能量转移。类似于福斯特共振能量转移(FRET),这种耦合显示出强烈的距离依赖,但在150纳米的范围内,并能够在1000光子的光子预算下以5-6纳米的分辨率对荧光团进行轴向定位。[3,4]在这里,我们发现使用石墨烯层,MIET的定位精度达到了埃的精度。在这样的精度下,微小的细节,如样品表面的纳米级粗糙度变得重要。为了证明原理,我们确定了具有先验已知的样品几何形状的样品从表面的单分子的绝对距离。我们在三种不同的衬底上旋转涂覆荧光染料分子(Atto655),间隔层厚度分别为10、15和20 nm,定义了分子与石墨烯层的距离。接下来,我们通过定位附着在脂质双分子层底部和顶部小叶的脂质头基团上的荧光染料来确定脂质双分子层(SLB)的厚度。我们已经证明,通过在MIET中使用石墨烯作为能量受体,在光子预算下的轴向定位精度和分辨率达到了传统SMLM实验中典型的亚纳米水平。石墨烯- miet的一个有趣的特点是,它提供了轴向定位精度,现在大大超过了大多数SMLM方法提供的横向定位精度。[1]A. I. Chizhik等。光子学报,2014,24 (2).[2]S. Isbaner等。生物工程学报,2018,35 (6).[3]N. Karedla等。化学工程学报,2014,37 (4).[4]N. Karedla等。j .化学。物理学报,2018,34(5):555 - 557。
Graphene-induced energy transfer: resolving distances at the Angström scale (Conference Presentation)
In recent years single-molecule localization super-resolution microscopy (SMLM) has become an indispensable tool for many fields of research. Here, for any image of a single molecule one determines its center position with much higher accuracy than the size of that image itself. A challenge of SMLM is to achieve super-resolution also along the third dimension. Recently, Metal-Induced Energy Transfer or MIET [1,2] was introduced. It exploits the energy transfer from an excited fluorophore to plasmons in a thin metal film. Similar to Forster Resonance Energy Transfer (FRET), this coupling shows a strong distance dependence, but over a range up to 150 nm and enables axial localization of fluorophores with 5-6 nm resolution at a photon budget of 1000 photons. [3,4] Here, we show that using a graphene layer the localization accuracy of MIET reaches Angstrom accuracy. At such accuracy, minute details such as nanometer-scale roughness of the sample surfaces becomes important.
For proof of principle, we determined absolute distances of single molecules from a surface for samples with an a priori well-known sample geometry. We spin-coated fluorescent dye molecules (Atto655) on top of three different substrates with spacer thickness values of 10, 15, and 20 nm, defining the distance of the molecules from the graphene layer. Next, we determined the thickness of supported lipid bilayers (SLBs) by localizing fluorescent dyes attached to lipid head groups in the bottom and top leaflet of the SLB.
We have demonstrated that by using graphene as the energy acceptor in MIET, the axial localization accuracy and resolution reaches sub-nanometer levels at photon budgets which are typical in conventional SMLM experiments. An interesting feature of graphene-MIET is that it provides an axial localization accuracy which now surpasses significantly that of lateral localization provided by most SMLM approaches.
[1] A. I. Chizhik et al. Nat. Photonics 8, 124 (2014).
[2] S. Isbaner et al. Nano Lett. 18, 2616 (2018).
[3] N. Karedla et al. ChemPhysChem 15, 705 7 (2014).
[4] N. Karedla et al. J. Chem. Phys. 148, 204201 (2018).