Tools For Measuring Free Zinc (pZn) In The Brain

描述（由申请人提供）：我们将为基础和临床神经科学家提供他们需要的工具来测量生物体液中的“游离”（快速交换）锌。事实证明，体内或体外脑组织的健康对游离锌浓度（pZn）和游离质子浓度（ph）一样敏感。我们已经开发出第一种实用的pZn计，一种荧光仪器，具有试管内和光纤探针测量方式。我们的客户将包括：(i)对突触和细胞内锌信号感兴趣的神经科学家，（ii）治疗中风、头部创伤、心脏骤停和癫痫患者的临床医生，锌现在被认为是损伤脑细胞的最严重的毒素之一，以及（iii）在体外培养或研究细胞的生物学家，他们想要监测（和控制）其培养基的pZn。总的来说，这些市场代表了大约80,000名客户，耗材每年约500美元（科学家）至每年约10,000美元（临床医生）。在第一阶段，我们建立了原型，荧光锌测量工具，我们用它们来建立生理pZn在大脑中的值（7.7 +/- 0.3）。在第二阶段，我们将利用我们对pZn生理价值的了解和我们对专有锌传感荧光分子的获取，并：(i)完成基于溶液的荧光锌计的开发，“调谐”以测量生理pZn， （ii）开发和优化带有锌感应荧光分子附着在壁上的试管，用于“半自动”pZn测定，（iii）为试管开发一种倏逝波照明，以便即使在“脏”的试管溶液中也可以进行荧光锌测定，（iv）将倏逝波+表面测定适应于光纤“锌引入”。(v)完善锌导入体的附着和传感化学，使其可用于中期（天-周）颅内pZn的原位记录。

DESCRIPTION (provided by applicant): We will deliver to basic and clinical neuroscientists the tools they need to measure "free" (rapidly- exchangeable) zinc in biological fluids. As it turns out, the health of brain tissue in vivo or in vitro is just as sensitive to the free zinc concentration, or "pZn," as it is to the free proton concentration, or "pH." We have developed the first, practical pZn Meter, a fluorimetric instrument, with both in-cuvette and optical fiber probe measuring modalities. Our customers will include: (i) neuroscientists interested in synaptic and intracellular zinc signaling, (ii) clinicians treating victims of stroke, head trauma, cardiac arrest, and seizures, where zinc is now understood to be one of the worst of the toxins injuring brain cells, and (iii), biologists growing or studying cells in vitro who want to monitor (and control) the pZn of their media. Collectively, these markets represent about 80,000 customers, with consumables of ~ $500/yr (scientists) to ~$10,000/yr (clinicians). In Phase I, we built prototype, fluorimetric zinc measurement tools, and we used them to establish the value for physiological pZn in the brain (7.7 +/- 0.3). In Phase II we shall capitalize on our knowledge of the physiological value of pZn and our access to proprietary zinc-sensing fluorescent molecules and: (i) complete development of a solution-based fluorimetric pZn Meter, "tuned" to measure physiological pZn, (ii) develop and optimize cuvettes with the zinc-sensing fluorescent molecules attached to the walls, for "semi-automatic" pZn determinations, (iii) develop an evanescent wave illumination for the cuvette, so that the fluorimetric zinc assays can be done even with "dirty" solutions in the cuvette, (iv) adapt the evanescent wave+ surface assay to an optical fiber "zintrode," (v) perfect the attachment and sensing chemistry on zintrodes so that they can be used for medium-term (days-weeks) in situ recording of intracranial pZn.