Optoinjection for efficient targeted delivery of a broad range of compounds and macromolecules into diverse cell types
Optoinjection for efficient targeted delivery of a broad range of compounds and macromolecules into diverse cell types
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DOI：
10.1117/1.2168148
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发表时间：
2006-01-01
影响因子：
3.5
通讯作者：
Koller, MR
中科院分区：
文献类型：
--
作者：
Clark, IB;Hanania, EG;Koller, MR
将化合物和大分子有效地递送到活细胞中在许多领域中是必不可少的，包括基础研究、应用药物发现和临床基因治疗。不幸的是，目前的递送方法，如阳离子脂质和电穿孔，受到可采用的大分子和细胞的类型、低效率和/或细胞毒性的限制。为了解决这些问题，开发了基于激光介导的通过光注射将大分子递送到细胞中的新方法。利用自动化高通量仪器，激光使能分析和处理（LEAPTM）系统来阐明和优化影响光注射效率和毒性的几个参数。采用直接细胞照射的技术（i.例如，以特定的细胞坐标为目标）和基于网格的照射（即，例如，没有定位单个细胞）都成功地开发了。通过这两种技术，确定了多次连续低辐射暴露比单次高辐射暴露产生更有利的结果。各种物质被有效地光注入到许多细胞类型中，包括离子、小分子、葡聚糖、siRNA（小干扰RNA）、质粒、蛋白质和半导体纳米晶体。值得注意的是，传统递送方法难治的细胞以较低的毒性被有效地光注射。我们建立了光注入的广泛实用性，并且是第一个以自动化，高通量的方式展示其实施的人。(c)2006年，美国摄影光学仪器工程师学会。
Efficient delivery of compounds and macromolecules into living cells is essential in many fields including basic research, applied drug discovery, and clinical gene therapy. Unfortunately, current delivery methods, such as cationic lipids and electroporation, are limited by the types of macromolecules and cells that can be employed, poor efficiency, and/ or cell toxicity. To address these issues, novel methods were developed based on laser- mediated delivery of macromolecules into cells through optoinjection. An automated high-throughput instrument, the laser- enabled analysis and processing (LEAP (TM)) system, was utilized to elucidate and optimize several parameters that influence optoinjection efficiency and toxicity. Techniques mploying direct cell irradiation (i. e., targeted to specific cell coordinates) and grid- based irradiation (i. e., without locating individual cells) were both successfully developed. With both techniques, it was determined that multiple, sequential low radiant exposures produced more favorable results than a single high radiant exposure. Various substances were efficiently optoinjected - including ions, small molecules, dextrans, siRNAs (small interfering RNAs), plasmids, proteins, and semiconductor nanocrystals - into numerous cell types. Notably, cells refractory to traditional delivery methods were efficiently optoinjected with lower toxicity. We establish the broad utility of optoinjection, and furthermore, are the first to demonstrate its implementation in an automated, high- throughput manner. (c) 2006 Society of Photo- Optical Instrumentation Engineers.