OPTICAL NANOSENSORS FOR SUBCELLULAR CHEMICAL IMAGING

我们的目标是生产光学纳米传感器， 细胞膜和细胞内过程的化学成像。 这些传感器将监测pH值，钙，镁，钠， 钾、氯、氧、亚硝酸盐、一氧化氮、二氧化碳 和葡萄糖。这些传感器所需的样本量为 比目前的光纤传感器少一百万到十亿倍， 绝对检测极限也是如此。 这些传感器还将 小100 - 1000倍，响应时间100 - 1000倍 短.该方法利用了一种已经证明的能力， 建造这种坚固的超小型传感器，并在一个新的 制备荧光阳离子和阴离子传感器的通用技术， 基于利用最好的离子载体， 电化学传感器具体而言，我们建议， 演示上述列表的亚微米光纤传感器 分析物。对于相同的分析物，我们还提出了一种新的，细胞 可植入光学纳米传感器（"卵石"），其是生物相容的 只占细胞体积的百万分之一。一群 细胞内卵石传感器可以提供多种分析物 信息或单个分析物通量。建议的一氧化氮 生物传感器是第一个将可逆性与联合收割机相结合的光学NO传感器， 选择性和快速响应，具有优异的检测限。这 设计也是可扩展的。二氧化碳镁离子 钙离子传感器也是一种新的设计和测量 活动 此外，刚刚构建了一个高级版本的 扫描近场光学显微镜，我们计划将其与联合收割机结合， 我们的光纤传感器变成一个扫描光纤化学图像 空间分辨率低至100 - 200 nm， 时间分辨率细胞和亚细胞化学成像将是 在大鼠胚胎、小鼠卵母细胞和人类胚胎上进行 神经母细胞瘤细胞这种纳米化学传感器和映射是 预计将彻底改变生物医学研究：由此产生的 技术将以不同的数量级加速， 生化测试协议，也使细胞 对亚细胞样品进行微生物学研究， 通道分辨率，导致化学时空映射作为 功能电生理细胞输入。长期目标是 实现单分子上的化学空间分辨率， 化学敏感性

The objective is to produce optical nanosensors for direct, real-time chemical imaging of cellular membranes and intracellular processes. These sensors will monitor pH, calcium, magnesium, sodium, potassium, chloride, oxygen, nitrite, nitric oxide, carbon dioxide and glucose. The sample size required for these sensors will be a million to a billion times less than for current fiber-optic sensors, and so will the absolute detection limit. These sensors will also be 1O0-1000 times smaller and their response times 1OO-1000 times shorter. The method capitalizes on an already demonstrated ability to construct such robust and ultra-small sensors and on a new universal technique for making fluorescent cation and anion sensors, based on utilizing as is the best ionophores available from electrochemical sensors. Specifically, we propose to make and demonstrate submicrometer fiber-optic sensors for the above list of analytes. For the same analytes we also propose a new, cell immplantable optical nanosensor ("pebble"), which is biocompatible and only occupies one millionth of the cell's volume. A group of intracellular pebble sensors can provide either multiple analyte information or single analyte fluxes. The proposed nitric oxide biosensor is the first optical NO sensor to combine reversibility, selectivity and fast response with an excellent detection limit. This design is also miniaturizable. The carbon dioxide, magnesium ion and calcium ion sensors are also new designs and measure activities. Also, having just constructed an advanced version of a scanning near-field-optical microscope, we plan to combine it with our fiber-optic sensors into a scanning-fiber-optic chemical imagery with a spatial resolution down to 100-200 nm and a millisecond time resolution. Cellular and subcellular chemical imaging will be carried out on rat embryos, mouse oocytes and human neuroblastoma cells. Such nano-chemical sensors and mappings are expected to revolutionize bio-medical research: the resulting technology will speed up by orders of magnitude various biochemical test protocols and also make possible cellular microbiology investigations on subcellular samples, with single channel resolution, resulting in chemical space-time mappings as a function electrophysiological cell input. The long range goal is to achieve single molecule on of the chemical spatial resolution and chemical sensitivity.