SPECTROGRAPHIC MULTIFLUOR LABELLED DIAGNOSTIC ARRAYS

目前的荧光标记诊断阵列阅读器（包括“基因芯片”）是激光扫描共聚焦显微镜的改编。它们具有较差的复用能力，并且不适合在高反射基板（如玻璃和硅）上成像网格。我们提出了一种基于新型成像光谱仪的荧光阵列读取器（FAR），能够同时激发和光谱分辨多种荧光，我们的初始目标是7个荧光（组合复杂度127'）。该设计将具有几微米的空间分辨率。对基材反射伪影有超强的免疫力。它非常适合成像弱荧光信号在强反射基板。并且将适应任何阵列格式，从具有毫米特征尺寸的粗糙手工点阵到包含10(6)个或更多元素cm-2的纳米阵列。光路不包含除试样台以外的任何移动部件，也不包含滤光片。并行多氟特征分析在一次通过中完成。全分辨率下的检测限约为100个氟分子μ m-1。读者的目标是“下一代”诊断阵列，其中组合标记方法将允许同时测试多个探针每个点的患者DNA，并分析来自多个患者的混合DNA使用相同的诊断阵列。拟议的商业应用：这项研究将开发一种自动读取器，用于临床和研究实验室使用的多荧光标记诊断阵列的高通量并行成像和分析。应用包括同步多基因分析，表达谱，高通量筛选和测序。

Current readers for fluor-labelled diagnostic arrays (including 'gene chips') are adaptations of laser-scanning confocal microscopes. They have poor multiplex capability' and are ill suited to imaging grids on high-reflectance substrates such as glass and silicon. We propose to develop a fluorescence array reader (FAR) based on a novel imaging spectrograph capable of exciting and spectrally resolving multiple fluors simultaneously our initial goal is 7 fluors (combinatorial complexity 127'). The design will have a spatial resolution of a few microns. and exceptional immunity from substrate reflectance artifacts. It is ideally suited to imaging weak fluorescent signals on strongly reflective substrates. and will accommodate any array format, from coarse manually-spotted grids with millimeter feature sizes down to nanofabricated arrays containing 10(6) or more elements cm-2. The optical path contains no moving parts except the specimen stage, and no filters. Parallel multi-fluor signature analysis is done in a single pass. Detection limit at full resolution will be on the order of 100 fluor molecules mu m-1. The reader is aimed at 'next generation' diagnostic arrays, in which combinatorial labelling methods will permit simultaneous testing with multiple probes per spot of patient DNA, and analysis of pooled DNA from multiple patients using the same diagnostic array. PROPOSED COMMERCIAL APPLICATIONS: This research will develop an automated reader for high throughput parallel imaging and analysis of multi-fluor labelled diagnostic arrays for clinical and research laboratory use. Applications include simultaneous multi-gene analysis, expression profiling, high-throughput screening, and sequencing.