Optical nanosensors for chemical analysis inside single living cells. 1. Fabrication, characterization, and methods for intracellular delivery of PEBBLE sensors
Optical nanosensors for chemical analysis inside single living cells. 1. Fabrication, characterization, and methods for intracellular delivery of PEBBLE sensors
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DOI：
10.1021/ac990629o
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发表时间：
1999-11-01
影响因子：
7.4
通讯作者：
Kopelman, R
中科院分区：
文献类型：
--
作者：
Clark, HA;Hoyer, M;Kopelman, R
球形光学纳米传感器，或鹅卵石（由生物定位嵌入封装的探针），已经生产出直径为20和200纳米的尺寸。这些传感器是在微乳液中制造的，由包裹在聚丙烯酰胺基质中的荧光指示器组成。一种广泛的聚合方法已经发展，允许生产传感器含有任何亲水性染料或染料的组合在基质中。PEBBLE基质保护荧光染料免受蛋白质的干扰，允许染料可靠的体内校准，传感器响应时间小于1ms。细胞活力测定表明，与对照条件相比，鹅卵石具有生物相容性，生物效应可以忽略不计。已经研究了几种传感器递送方法，包括脂质体递送、基因枪轰击和皮注射到单个活细胞中。
Spherical optical nanosensors, or PEBBLEs (probes encapsulated by biologically localized embedding), have been produced in sizes including 20 and 200 nm in diameter. These sensors are fabricated in a microemulsion and consist of fluorescent indicators entrapped in a polyacrylamide matrix. A generalized polymerization method has been developed that permits production of sensors containing any hydrophilic dye or combination of dyes in the matrix. The PEBBLE matrix protects the fluorescent dye from interference by proteins, allowing reliable in vivo calibrations of dyes, Sensor response times are less than 1 ms. Cell viability assays indicate that the PEBBLEs are biocompatible, with negligible biological effects compared to control conditions. Several sensor delivery methods have been studied, including liposomal delivery, gene gun bombardment, and picoinjection into single living cells.