BRIDGE C--LC/Mass Spectrometric Analysis/Neutral Lipids

BRIDGE C--LC/质谱分析/中性脂质

基本信息

批准号：
7551345
负责人：
ROBERT Carl MURPHY
金额：
$ 25.49万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/7551345
关键词：
Animals Aorta Area Arterial Fatty Streak Atherosclerosis Award Basic Science Biopsy Cells Cellular biology Chemistry Classification Compatible Coupled Data Collection Data Set Databases Deposition Detection Development Diabetes Mellitus Diffusion Dimensions Disease Generations Goals Image Ions Lasers Lateral Lipid Peroxidation Lipids Location Microdissection Modification Perfusion Phase Phospholipids Positioning Attribute Process Rate Relative (related person)Reproducibility Research Research Methodology Resolution Scanning Shapes Signal Transduction Slice Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Spectrometry, Mass, Secondary Ion Sphingolipids Sublimation - mental defence mechanism Techniques Time Tissue Embedding Tissue Sample Tissues base fullerene C60 human subject improved interest irradiation mass spectrometer novel programs research study size software development success vapor

项目摘要

Bridge C will carry out basic research with the objective to develop mass spectrometric techniques that can be used to generate images of the specific location and relative abundance of lipids in tissues and cells. In part, this research program builds on the success during the first award period of using matrix assisted laser desorption ionization (MALDI) coupled to a quadrupole tandem time-of-flight mass spectrometer and application of matrix by sublimation onto thin tissues slices (10 um) to enhance ion yields following irradiation of small areas of tissue (approximately 100 um2) with a focused laser beam. The resultant secondary ions emitted are then mass analyzed and stored as the mass and intensity values at that pixel. The adjacent areas are irradiated sequentially to build a large database that can be used to generate images specific to the lipid ion (m/z) as a function of position and ion abundance. Sublimation of matrix was found to minimize lipid lateral diffusion in tissues and allow generation of detailed images of tissues based upon mass spectrometric pixels. Secondary ion mass spectrometry (SIMS) using Buckminster fullerene (C60 +) ion beams will be systematically studied to bring lateral resolution of lipid imaging below 1 um so that lipids in single cells can be detected and analyzed. Specific goals include development of derivatization chemistry to enhance lipid secondary ion yield in MALDI and SIMS imaging experiments to allow discovery of novel lipids. New tissue embedding materials will be developed and explored that will be specifically suited for mass spectrometric imaging. The reproducibility of lipid anatomical images will be assessed by analysis of sequential tissue slices as well as of identical tissue regions from different animals. Threedimensional rendering of lipid images from sequential slices will be pursued using software developed for MALDI and SIMs data sets. A major focus will be imaging of aortas and atherosclerotic plaques as part of the LIPID MAPS coordinated studies to reveal anatomical locations of specific lipids including but not limited to phospholipids, sphingolipids, and oxidized neutral lipids. In addition, this research will develop novel and powerful research methods that can be used to examine tissues (biopsies) taken from human subjects for molecules that mark unique disease processes such as atherosclerosis and diabetes.

Bridge C 将开展基础研究，目标是开发质谱技术可用于生成组织和细胞中脂质的特定位置和相对丰度的图像。在某种程度上，该研究计划建立在使用矩阵辅助的第一个奖励期的成功基础上激光解吸电离 (MALDI) 与四极串联飞行时间质谱仪耦合通过升华将基质应用到薄组织切片（10 um）上以提高离子产量用聚焦激光束照射小区域的组织（大约 100 um2）。由此产生的然后对发射的二次离子进行质量分析并将其存储为该像素处的质量和强度值。相邻区域依次受到照射，建立一个大数据库，可用于生成特定于脂质离子 (m/z) 的图像，作为位置和离子丰度的函数。基质的升华是发现可以最大限度地减少组织中的脂质横向扩散，并允许生成基于组织的详细图像基于质谱像素。使用巴克明斯特富勒烯的二次离子质谱 (SIMS) (C60 +) 将系统地研究离子束，使脂质成像的横向分辨率低于 1 um，以便可以检测和分析单细胞中的脂质。具体目标包括衍生化的发展化学方法可提高 MALDI 和 SIMS 成像实验中的脂质二次离子产量，从而实现发现新型脂质。将开发和探索新的组织包埋材料，专门用于适用于质谱成像。脂质解剖图像的再现性将通过以下方式进行评估分析连续组织切片以及来自不同动物的相同组织区域。三维将使用为以下目的开发的软件来渲染连续切片的脂质图像 MALDI 和 SIM 数据集。主要重点是主动脉和动脉粥样硬化斑块的成像，作为 LIPID MAPS 协调研究以揭示特定脂质的解剖位置，包括但不限于磷脂、鞘脂和氧化中性脂质。此外，这项研究将开发新颖且强大的研究方法，可用于检查从人类受试者身上获取的组织（活检）标记动脉粥样硬化和糖尿病等独特疾病过程的分子。