Tools For Measuring Free Zinc (pZn) In The Brain

描述（由申请人提供）：我们将向基础和临床神经科学家提供他们测量生物液体中“游离”（快速交换）锌所需的工具。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值）同样敏感。“我们已经开发出第一台实用的pZn Meter，这是一种荧光仪器，具有比色皿内和光纤探头测量模式。我们的客户包括：（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，心脏骤停和癫痫发作患者的临床医生，其中锌现在被认为是伤害脑细胞的最严重的毒素之一，以及（iii）想要监测（和控制）其介质的pZn的体外培养或研究细胞的生物学家。总的来说，这些市场代表了大约80，000个客户，消耗品约为500美元/年（科学家）至10，000美元/年（临床医生）。在第一阶段，我们建立了原型，荧光锌测量工具，我们用它们来建立大脑中生理pZn的值（7.7 +/- 0.3）。在第二阶段，我们将利用我们对pZn的生理价值的了解以及我们对专有锌传感荧光分子的访问，并且：（i）完成基于溶液的荧光pZn计的开发，“调谐”以测量生理pZn，（ii）开发和优化具有附接到壁的锌感测荧光分子的比色皿，用于“半自动”pZn测定，（iii）开发用于比色皿的渐逝波照明，使得即使在比色皿中存在“脏”溶液的情况下也可以进行荧光锌测定，（iv）使渐逝波+表面测定适应于光纤“zintrode”，“（v）完善锌电极上的附着和传感化学，以便它们可以用于颅内pZn的中期（数天-数周）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.