Engineering Development of Laser Capture Microdissection

激光捕获显微切割是美国国立卫生研究院开发的一项技术，可以从组织病理切片中捕获微小的组织部分，用于后续的分子分析。与组织样本接触的热塑性薄膜在显微镜控制下被激光选择性地激活，以与所需组织结合。去除薄膜会将附着的组织与载玻片上剩余的组织分开，使其可用于分析。这项技术能够以常规方式收集和处理极其均一的样品，而不需要很大的手工灵活性。在过去的几年里，对这项技术进行了许多改进。我们已经完善了圆柱形结构，它采用了与组织的“线”接触，以将污染降至最低。这能够去除直径小至3微米的组织碎片。我们进一步将其发展为非接触式系统，以优化单细胞传输系统。我们还开发了一种非接触式系统，利用一条连续的热塑性胶带，与组织之间有大约5微米的间隙。胶带和组织之间的接触将仅在激光脉冲的直接位置发生。我们正在探索使用这种替代基板是否具有性能和经济优势。

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 have also developed a non-contact system utilizing a continuous tape of thermoplastic which is separated from the tissue by a gap of approximately 5 microns. Contact between the tape and the tissue will only occur at the immediate location of the laser pulse. We are exploring whether there are performance and economic advantages to using this alternative substrate.