OPTICAL NANOSENSORS FOR SUBCELLULAR CHEMICAL IMAGING

其目标是生产用于直接、实时的光学纳米传感器 细胞膜和细胞内突起的化学成像。 这些传感器将监测pH、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大小将为 比目前的光纤传感器少100万到10亿倍， 绝对探测极限也是如此。这些传感器也将是 100-1000倍小，响应时间100-1000倍 再短一点。这种方法充分利用了已经证明的能力。 为了构建如此坚固和超小型的传感器，并建立在一种新的 制造阳离子和阴离子荧光传感器的通用技术， 基于对As的利用，是可从 电化学传感器。具体地说，我们建议制定和 演示上述列表中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 植入生物兼容的光学纳米传感器(“鹅卵石”) 只占细胞体积的百万分之一。一群 细胞内的卵石传感器可以提供多种分析物 信息或单一分析物通量。拟议中的一氧化氮 生物传感器是第一个结合可逆性的光学NO传感器， 选择性好，响应快，检测下限好。这 设计也是可以微型化的。二氧化碳、镁离子 而钙离子传感器也是新的设计和测量 活动。此外，在刚刚构建了高级版本的 扫描近场光学显微镜，我们计划将其与 我们的光纤传感器变成了扫描光纤化学图像 空间分辨率降至100-200 nm和1毫秒 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身上进行了研究 神经母细胞瘤细胞。这种纳米化学传感器和映射是 预计将给生物医学研究带来革命性的变化：由此 技术将以不同数量级的速度 生化测试协议，并使细胞 亚细胞样品的微生物学研究 通道分辨率，导致化学时空映射作为 功能电生理细胞输入。远景目标是 实现了单分子的化学空间分辨率和 化学敏感度。

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.