3D printed microchannels for sub-nL NMR spectroscopy.
3D printed microchannels for sub-nL NMR spectroscopy.
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DOI：
10.1371/journal.pone.0192780
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发表时间：
2018
期刊：
影响因子：
3.7
通讯作者：
Boero G
中科院分区：
文献类型：
--
作者：
Montinaro E;Grisi M;Letizia MC;Pethö L;Gijs MAM;Guidetti R;Michler J;Brugger J;Boero G
亚纳升(亚纳米升)体积的核磁共振实验由于探测器的有限灵敏度以及在探测器附近定位和保持如此小的样品的困难而受到阻碍。在这项工作中，我们报道了液体和生物实体在体积小于100pl的液体中的核磁共振实验。这些测量是通过制造高空间分辨率3D打印微流控结构实现的，该结构专门用于引导和限制亚NL样品在单芯片集成核磁共振探头的亚NL最灵敏体积中。微流体结构是使用分辨率优于1μm~3的双光子聚合3D打印技术制造的。这里采用的高空间分辨率3D打印方法允许快速制造复杂的微流控结构，以定位、保持和进样生物样品，其设计最大限度地提高了核磁共振信号的幅度，并将静态磁场的不均匀降至最低。将样品与微线圈分开的层厚度为10μm，这对于发挥探测器的最高灵敏度至关重要。为了展示这种方法的潜力，我们报告了液体介质中亚NL完整生物实体的核磁共振实验，特别是迟发性里氏冠状线虫的卵子和线虫的切片。我们在7T时对1H核的灵敏度为2.5×1013自旋/Hz1/2，足以在3小时的测量时间内检测到活性体积降至100pl的内源化合物的1H核的6pmoL。在液体样品中的光谱分辨率为0.01ppm，在所研究的生物实体中的光谱分辨率为0.1ppm。所获得的结果可能为在单个单位水平上进行具有亚NL体积的重要生物实体的核磁共振研究指明了路线，例如包括人类在内的几种哺乳动物的活的微生物和卵。
Nuclear magnetic resonance (NMR) experiments on subnanoliter (sub-nL) volumes are hindered by the limited sensitivity of the detector and the difficulties in positioning and holding such small samples in proximity of the detector. In this work, we report on NMR experiments on liquid and biological entities immersed in liquids having volumes down to 100 pL. These measurements are enabled by the fabrication of high spatial resolution 3D printed microfluidic structures, specifically conceived to guide and confine sub-nL samples in the sub-nL most sensitive volume of a single-chip integrated NMR probe. The microfluidic structures are fabricated using a two-photon polymerization 3D printing technique having a resolution better than 1 μm3. The high spatial resolution 3D printing approach adopted here allows to rapidly fabricate complex microfluidic structures tailored to position, hold, and feed biological samples, with a design that maximizes the NMR signals amplitude and minimizes the static magnetic field inhomogeneities. The layer separating the sample from the microcoil, crucial to exploit the volume of maximum sensitivity of the detector, has a thickness of 10 μm. To demonstrate the potential of this approach, we report NMR experiments on sub-nL intact biological entities in liquid media, specifically ova of the tardigrade Richtersius coronifer and sections of Caenorhabditis elegans nematodes. We show a sensitivity of 2.5x1013 spins/Hz1/2 on 1H nuclei at 7 T, sufficient to detect 6 pmol of 1H nuclei of endogenous compounds in active volumes down to 100 pL and in a measurement time of 3 hours. Spectral resolutions of 0.01 ppm in liquid samples and of 0.1 ppm in the investigated biological entities are also demonstrated. The obtained results may indicate a route for NMR studies at the single unit level of important biological entities having sub-nL volumes, such as living microscopic organisms and eggs of several mammalians, humans included.
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3D打印的微流体。
DOI：
10.1002/anie.201504382
发表时间：
2016-03-14
期刊：
Angewandte Chemie (International ed. in English)
影响因子：
--
作者：
Au AK;Huynh W;Horowitz LF;Folch A
通讯作者：
Folch A
影响因子：
2.2
作者：
Anders, Jens;Chiaramonte, Giuseppe;Boero, Giovanni
通讯作者：
Boero, Giovanni
影响因子：
2.2
作者：
Finch, Graeme;Yilmaz, Ali;Utz, Marcel
通讯作者：
Utz, Marcel
影响因子：
64.8
作者：
AGUAYO, JB;BLACKBAND, SJ;HINTERMANN, M
通讯作者：
HINTERMANN, M
影响因子：
3.3
作者：
Grant, SC;Buckley, DL;Blackband, SJ
通讯作者：
Blackband, SJ