Monolithic Silica Microbands Enable Thin-Layer Chromatography Analysis of Single Cells.
Monolithic Silica Microbands Enable Thin-Layer Chromatography Analysis of Single Cells.
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DOI：
10.1021/acs.analchem.2c02622
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发表时间：
2022-10-04
影响因子：
7.4
通讯作者：
Allbritton, Nancy L.
中科院分区：
文献类型：
--
作者：
Wang, Yuli;Yao, Ming;Sims, Christopher E.;Allbritton, Nancy L.
皮升薄层色谱（pTLC）平台被开发用于分析极其微小的样品，例如测定1皮升体积的单个细胞的内容物。pTLC芯片由设计用于接受皮升级体积样品的高度多孔的单片二氧化硅制成的微尺度带阵列组成。结合溶胶-凝胶化学和微加工技术制备pTLC谱带。每个微带的宽度（60-80 μm）和深度（13 μm）与单个细胞的大小相当，并起到减少侧向扩散和限制化合物沿着微带运动的作用。通过压电微分配器将超小体积（数十pL）的模型荧光化合物点样到微带上，并通过pTLC成功分离。在溶胶-凝胶过程中，通过改变致孔剂的浓度可以调节大孔的大小（0.3 ~ 2.3 μm），从而提高了分离分辨率和分析物的迁移率。对于0.3 μm大孔径，使用标准显微镜方法通过荧光可检测到阿摩尔分析物。分离分离度、理论塔板数和分离时间分别为1.3-2.1、4-357和2-8 min。为了证明pTLC从单个哺乳动物细胞中分离分析物的能力，将加载有荧光亲脂性染料或鞘氨醇激酶报告基因的细胞点样在微带上，并从其荧光检测通过pTLC分离的单细胞内容物。这些结果表明pTLC在许多需要微型样品和高通量平行分析的领域中的应用潜力。
A picoliter thin layer chromatography (pTLC) platform was developed for analyzing extremely miniature specimens, such as assay of the contents of a single cell of 1 picoliter volume. The pTLC chip consisted of an array of microscale bands made from highly porous monolithic silica designed to accept picoliter-scale volume samples. pTLC bands were fabricated by combining sol-gel chemistry and microfabrication technology. The width (60–80 μm) and depth (13 μm) of each band is comparable to the size of single cells and acted to reduce the lateral diffusion and confine the movement of compounds along the microbands. Ultra-small volumes (tens of pL) of model fluorescent compounds were spotted onto the microband by a piezoelectric microdispenser and successfully separated by pTLC. The separation resolution and analyte migration were dependent on the macropore size (ranging from 0.3 to 2.3 μm) which was adjustable by changing the porogen concentration during the sol-gel process. For a 0.3 μm macropore size, attomoles of analyte were detectable by fluorescence using standard microscopy methods. The separation resolution, theoretical plate number and separation times ranged from 1.3–2.1, 4–357, and 2–8 min, respectively for the chosen model biological lipids. To demonstrate the capability of pTLC for separating analytes from single mammalian cells, cells loaded with fluorescent lipophilic dyes or sphingosine kinase reporter were spotted on microbands, and the single-cell contents separated by pTLC were detected from their fluorescence. These results demonstrate the potential of pTLC for applications in many areas where miniature specimens and high throughput parallel analyses are needed.
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影响因子：
7.4
作者：
Bakry, Rania;Bonn, Gunther K.;Svec, Frantisek
通讯作者：
Svec, Frantisek
影响因子：
3.2
作者：
Kim, Young Kwon;Park, Ju An;Jung, Sungjune
通讯作者：
Jung, Sungjune
影响因子：
4.1
作者：
Schnute, Mark E.;McReynolds, Matthew D.;Nagiec, Marek M.
通讯作者：
Nagiec, Marek M.
影响因子：
1.6
作者：
Hauck, HE;Bund, O;Schulz, M
通讯作者：
Schulz, M
DOI：
10.1016/b978-0-12-811732-3.00005-4
发表时间：
2017-01-01
期刊：
HANDBOOK OF ADVANCED CHROMATOGRAPHY/MASS SPECTROMETRY TECHNIQUES
影响因子：
--
作者：
Svec, Frantisek;Lv, Yongqin
通讯作者：
Lv, Yongqin