DEVELOPMENT OF AN INTELLIGENT SAMPLE INTRODUCTION SYSTEM FOR MASS SPECTROMETRY

产品说明：质谱智能进样系统的研制迫切需要新的技术，使快速测量，而不显着妥协的分析数据。在此，我们建议开发一种基于质谱（MS）的表面采样装置，该装置能够直接分析各种材料，包括生物组织和活微生物，从而实现这一目标。该装置的基础是液体微结表面取样探针（LMJSSP），这是一组同轴微流体毛细管，其被泵送和抽吸以将溶剂带到和远离感兴趣的表面，其中在探针和样品之间的界面处发生微萃取。Prosolia已从AB Sciex获得LMJSSP技术的许可，并将通过NIH实验室到市场计划与橡树岭国家实验室的原始发明者合作，通过使该技术在各种生物分析工作流程中的应用更加强大和用户友好，为商业上可行的产品创建先进的原型。该项目的任务可以分为三个目标。第一和第二目的是确保设备的硬件足够容易使用， 商业可行性。这包括表征本原型和探索的几个有前途的方法，自动控制匹配的液体流量所需的连续提取交界处探针和表面之间持续。此外，建立一个机制，保持探头和表面之间的理想距离将是成功实施的关键。在建立具有优化控制和合理生产潜力的流量和高度管理方法后，这些硬件修改将用于确定仪器操作的分析条件。具体而言，将创建标准，以确定在各种条件下（包括在不同实验室内）的提取概况和效率。这些将确定潜在使用、维护和支持的预期限制，以确保针对各种方法校准的稳健操作。在本部分工作中，将通过较小的变体对标准探测布置的分辨率的潜在增加进行调查，以确定诊断和生物标志物研究人员感兴趣的组织学相关样本量的分析指标。早期实验使用另一种基于MS的表面采样技术，解吸电喷雾电离，用于活微生物的直接生物分析，能够生成细菌菌落的光谱曲线;然而，由于细菌从表面吹到仪器入口的污垢，产生这些结果的过程需要频繁清洁和更换仪器毛细管。使用我们的原型LMJSSP系统获得的初步光谱已经收集在一个更容易的方式，并已显示出一些特别有趣的功能，用于区分和表征不同的物种和菌株的细菌。然而，必须获得更多的指纹，以确定这些数据的统计学意义和适用性。此外，我们将建立在各种生长点和强制降解条件下的取样效果，以生成用于自动分类的潜在文库。在实现自动化和优化的变化后，我们用于表面直接分析的原型将是一个简单的样品引入系统，从少数人使用的学术原型过渡到可以部署到许多人的商业原型。通过遵循我们提出的里程碑：进行适当的硬件变更，确定与溶剂和探针大小相对应的分析指标，并收集统计学相关数量的细菌分析概况，将评估手动微生物分析的能力。因此，我们的第二阶段提案将包括进一步改进硬件、更广泛的样本选择、增强的数据集成和处理软件。

DESCRIPTION: Development of an Intelligent Sample Introduction System for Mass Spectrometry There is a pressing need for new technologies that enable rapid measurements without significant compromise to the analytical data. Herein, we propose to develop a robust mass spectrometry (MS) based surface sampling device that enables direct analysis of a variety of materials including biological tissue and live microorganisms that promises to deliver on this objective. The basis for this device is the Liquid Micro junction Surface Sampling Probe (LMJSSP), a set of coaxial microfluidic capillaries that are pumped and aspirated to bring solvent to and away from a surface of interest, where micro extraction takes place at the interface between the probe and the sample. Prosolia has licensed the LMJSSP technology from AB Sciex and will partner with the original inventor at Oak Ridge National Laboratory via the NIH Lab to Marketplace program to create an advanced prototype towards a commercially viable product by making the technology more robust and user-friendly for applications in various bio analytical workflows. The tasks of this project can be divided into three Aims. The first and second Aims are to ensure that the hardware of the device is sufficiently easy to use for commercial viability. This includes characterization of the present prototype and exploration of a few promising methods for automatic control of matched liquid flows required for a continuous extractive junction to persist between probe and surface. Additionally, establishing a mechanism for maintaining an ideal distance between the probe and surface will be critical for successful implementation. Upon establishing methods for flow and height management with optimized control and reasonable potential for manufacture, these hardware modifications will be used to determine the analytical conditions for instrument operation. Specifically, standards will be created to determine the profile and efficiency of extraction under a variety of conditions (including within different laboratories). These will determine the expected limits for potential use, maintenance, and support to ensure robust operations calibrated for a variety of methodologies. During this section of work, investigations into potential increases in resolution from the standard probing arrangement via smaller variants will be performed to identify analytical metrics at histologically relevant sampling sizes of interest to diagnostic and biomarker researchers. Early experiments using another MS-based surface sampling technique, Desorption Electrospray Ionization, for direct bio analysis of live microorganisms were able to generate spectral profiles for bacterial colonies; however, the process to generate these results required frequent cleaning and exchanging of instrument capillaries due to fouling with bacteria that were blown off the surface and into the instrument inlet. Preliminary spectra obtained using our prototype LMJSSP system have been collected in a much more facile manner, and have shown some particularly interesting features for distinguishing and characterizing different species and strains of bacteria. However, more fingerprints must be acquired in order to determine statistical significance and applicability of these data. In addition, we will establish he effects of sampling at various growth points and stress conditions in order to generate a potential library for automated classification. After implementing changes for automation and optimization, our prototype for direct analysis of surfaces, will be a simple sample introduction system transitioning from an academic prototype used by a few to a commercial prototype that can be deployed to many. By hewing to the milestones we propose: making appropriate hardware changes, determining analytical metrics corresponding to solvents and probe sizes, and gathering a statistically relevant number of profiles for bacterial analysis, the capacity for hands-off microorganism profiling will have been assessed. As such our phase II proposal will include further hardware refinements, a broader selection of samples, enhanced data integration, and processing software.