A VERSATILE STANDARD FOR QUANTITATIVE LIGHT MICROSCOPY

定量光学显微镜是研究细胞功能的一种有价值的技术。通过定量测量外源和内源光学探针的光，获得了关于细胞内蛋白质相互作用、离子浓度、膜电位和其他形式的细胞活动的新数据。这些测量的准确性在很大程度上依赖于经过良好校准和表征的显微镜光检测系统。本提案描述了定量光学显微镜通用校准标准的发展。可靠且易于使用的高精度标准在商业上是不可获得的，这将极大地促进可重复性测量。目前用作显微镜光标准的技术存在一些实际困难。这些设备的校准光输出受到照明灯噪声和老化的影响。本方案中描述的电子标准的光输出与显微镜的照明器输出无关。它从单个光圈提供多个用户可选波长和照度级别的可重复平场照明。此外，该仪器还可以用来监测和记录显微镜照明器的强度水平。该仪器具有独特的多功能性，具有任何竞争产品所不具备的操作模式。这项技术代表了光学显微镜领域的一项重大技术创新。建议的商业应用：本建议中描述的通用电子标准代表了一种校准光学显微镜设备的新方法。与中性密度滤光片或荧光珠等竞争技术相比，它具有显著的优势。由于这些优势，这项技术可以在价值5000万美元的显微镜校准配件市场占据相当大的份额。

Quantitative light microscopy is a valuable technique for studies of cellular function. New data concerning interactions of intracellular proteins, ion concentrations, membrane potential and additional forms of cellular activity are obtained from quantitative measurement of light from exogenous and endogenous optical probes. The accuracy of these measurements relies heavily on well calibrated and characterized microscope light detection systems. This proposal describes the development of a versatile calibration standard for quantitative light microscopy. A reliable and easily used high precision standard is not commercially available and would greatly facilitate reproducible measurements. Technologies currently used as light standards for microscopy present some practical difficulties. The calibrated light output of these devices is affected by illuminator lamp noise and aging. The light output of the electronic standard described in this proposal is independent of the microscope's illuminator output. It provides repeatable flat-field illumination at multiple user selectable wavelengths and illumination levels from a single aperture. Additionally, the instrument can be used to monitor and record the intensity level of the microscope's illuminator. The instrument is uniquely versatile, having operating modes unavailable in any competing product. This technology represents a significant technical innovation in the field of light microscopy. PROPOSED COMMERCIAL APPLICATIONS: The versatile electronic standard described in this proposal represents a new approach for the calibration of light microscopy equipment. It has significant advantages compared to competing technologies, such as neutral density filters or fluorescent beads. Because of these advantages, this technology could capture a significant portion of the $50 million microscope calibration accessory market.