Microstamp patterns of biomolecules for high-resolution neuronal networks
Microstamp patterns of biomolecules for high-resolution neuronal networks
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DOI：
10.1007/bf02522871
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发表时间：
1998-01-01
影响因子：
3.2
通讯作者：
Wheeler, BC
中科院分区：
文献类型：
--
作者：
Branch, DW;Corey, JM;Wheeler, BC
为了定位神经元及其轴突和树突，已经开发了一种用于在玻璃衬底上高分辨率绘制蛋白质图案的微印花技术。图案化工艺使用微米长的聚二甲基硅氧烷印章，将多种类型的生物分子转移到硅烷衍生的底物上，并使用戊二醛作为同源双功能连接剂。为了测试该过程的有效性，比较了光致抗蚀剂物理吸附和图案化聚-D-赖氨酸(PDL)的衬底和压印了PDL的衬底。用异硫氰酸荧光素标记的聚L赖氨酸来验证玻璃基板上图案的存在和均匀。作为生物学检测，B104神经母细胞瘤细胞被接种在冲压和物理吸附的玻璃盖玻片上。以电镀8h后图案上细胞的百分比来确定图案的顺应性。结果表明，冲压工艺与光刻胶构图工艺是等价的。标记了PDL的衬底的平均图案符合率为52.6+/-4.4%，而物理吸附的衬底的图案符合率为54.6+/-8.1%。背景回避措施也是相同的。由于该程序允许连续标记多个蛋白质，每个蛋白质都有自己的微图案，因此它对于定义复杂的底物以帮助细胞构图和其他细胞指导研究应该是非常有用的。
A microstamping technique has been developed for high-resolution patterning of proteins on glass substrates for the localisation of neurons and their axons and dendrites. The patterning process uses a microfabricated polydimethylsiloxane stamp with micrometer length features to transfer multiple types of biomolecules to silane-derivatised substrates, using glutaraldehyde as a homobifunctional linker. To test the efficacy of the procedure, substrates are compared in which poly-d-lysine (PDL) was physisorbed and patterned by photoresist with those stamped with PDL. Fluorescein isothiocyanate labelled poly-l-lysine was used to verify the presence and uniformity of the patterns on the glass substrates. As a biological assay, B104 neuroblastoma cells were plated on stamped and physisorbed glass coverslips. Pattern compliance was determined as the percentage of cells on the pattern 8h after plating. Results indicate that the stamping and photoresist patterning procedure are equivalent. Substrates stamped with PDL had an average pattern compliance of 52.6+/-4.4%, compared to 54.6+/-8.1% for physisorbed substrates. Measures of background avoidance were also equivalent. As the procedure permits successive stamping of multiple proteins, each with its own micropattern, it should be very useful for defining complex substrates to assist in cell patterning and other cell guidance studies.