ENGINEERING DEVELOPMENT OF LASER CAPTURE MICRODISSECTION

激光捕获显微解剖是美国国立卫生研究院开发的一种技术，用于从组织病理切片中捕获组织的微小部分，用于随后的分子分析。在显微控制下，用激光束选择性地激活与组织试样接触的热塑性薄膜，使其与所需组织结合。除去薄膜将粘附组织与载玻片上的剩余组织分离，使其可用于分析。该技术能够以常规方式收集和处理极其均匀的样品，不需要很大的手工灵活性。在过去的几年里，这项技术得到了许多改进。我们已经完善了一个圆柱形结构，它采用与组织的直线接触来减少污染。它能够去除直径小至3微米的组织碎片。我们进一步将其发展为非接触式系统，以优化单细胞转移系统。我们还在开发一种非接触式系统，利用一种连续的热塑性胶带，该胶带与组织隔开约5微米的间隙。胶带和组织之间的接触只会发生在激光脉冲的直接位置。我们希望探索使用这种替代基板是否具有性能和经济优势。我们正在完善描述激光能量在聚合物中被吸收和扩散过程的热模型，以便进一步预测组织和薄膜温度瞬态作为系统参数的函数。-显微解剖，激光-捕获-显微解剖，组织-显微解剖，激光

Laser Capture Microdissection is a technique developed at NIH to capture minute portions of tissue from histo-pathology slides for subsequent molecular analysis. A thermoplastic film in contact with a tissue specimen is selectively activated by a laser beam under microscopic control to bond with the desired tissue. Removal of the film separates the adhered tissue from that remaining on the slide, making it available for analysis. The technique enables the collection and processing of extremely homogeneous samples in a routine manner that does not require great manual dexterity. Over the past several years a number of refinements of the technique have been developed. We have perfected a cylindrical configuration which employs a line contact with the tissue to minimize contamination. This is capable of removing tissue fragments as small as 3 microns in diameter. We have further developed this into a non-contact system to optimize the system for single cell transfers. We are also developing a non-contact system utilizing a continuous tape of thermoplastic which is separated from the tissue by a gap of approximatly 5 microns. Contact between the tape and the tissue will only occur at the immediate location of the laser pulse. We hope to explore whether there are performance and economic advantages to using this alternative substrate. We are refining our thermal model which describes the process by which the laser energy is absorbed and diffused in the polymer in order to further predict tissue and film temperature transients as a function of the system parameters. - microdissection, laser- capture- microdissection, tissue- microdissection, lasers