Bead-based cellular analysis, sorting and multiplexing

Bead-based cellular analysis, sorting and multiplexing
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DOI:
10.1002/cbic.200500059
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发表时间:
2005-08-01
期刊:
影响因子:
3.2
通讯作者:
Bradley, M
Bradley, M
中科院分区:
生物学3区
文献类型:
--
作者:
Sanchez-Martin, RM;Muzerelle, M;Bradley, M

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细胞进入对于潜在药物、小分子调节剂和传感器的递送非常重要。最近,人们的注意力集中在开发各种特定的递送系统,以增强细胞水平的运输和摄取。当代的递送载体包括肽,如基于Tat蛋白衍生片段的肽,拟肽载体,如基于聚合物和树状大分子的肽,以及各种物理方法,如微注射。[1-7]聚合物介导的递送具有许多优点;例如,聚合物材料可以在可控的局部浓度下控制其货物的输送和释放,并且有许多将药物和传感器封装在聚合物微粒中的例子。然而,它们进入细胞往往是复杂的一种已被证明可以被吞噬并在细胞内积极运输的聚合物材料是乳胶珠;然而,令人惊讶的是,很少有研究利用这一现象的工作其中一个原因是,商业上可用的材料通常在化学上是非功能化的,或者如果它们被功能化,这只能在温和的水条件下实现,而为了附着一系列更有趣的化学探针而进行的改性需要与有机溶剂的相容性。本研究的目的是产生具有一定尺寸的稳定的生物相容性聚合物颗粒,这些颗粒可以用于固相多步骤合成,并且可以迅速被细胞吸收并用于各种应用。本文描述了一系列功能化、交联、多标记的聚合物微球作为穿越活细胞膜的一种模式的应用。它允许一般细胞标记,分类和动态细胞内过程的潜在监测。
Cellular entry is important for the delivery of potential drugs, small-molecule modulators and sensors. Recently attention has focused on the development of a diverse range of specific delivery systems in order to enhance transport and uptake at the cellular level. Contemporary examples of delivery vectors include peptides, such as those based on fragments derived from the Tat protein, peptidomimetic carriers, such as those based on polymers and dendrimers, and various physical methods, such as micro-injection.[1–7] Polymer-mediated delivery offers many advantages; for instance, polymeric materials can allow the controlled delivery and release of their cargo at controllable local concentrations, and there are many examples of drugs and sensors being encapsulated within polymer microparticles. However, their introduction into cells is often complex.[7] One type of polymeric material that has been shown to be engulfed and actively transported throughout cells is latex beads; however, surprisingly little work has been carried out on exploiting this phenomenon.[8] One reason for this is that commercially available materials are often unfunctionalized chemically or, if they are functionalized, this is only achievable under mild aqueous conditions, whereas modification for the attachment of a range of more interesting chemical probes requires compatibility with organic solvents.The aim of this study was the generation of robust biocompatible polymeric particles of defined sizes that could be employed in solid-phase multistep syntheses yet be taken up rapidly by cells and exploited in a variety of applications. Here the application of a range of functionalized, cross-linked, multilabelled polymer microspheres as a mode of traversing the membrane of living cells is described. It allows general cellular labelling, sorting and the potential monitoring of dynamic intracellular processes.