Tangrea Michael A, Hanson Jeffrey C, Bonner Robert F, Pohida Thomas J, Rodriguez-Canales Jaime, Emmert-Buck Michael R
Pathogenetics Unit and Laser Microdissection Core, Laboratory of Pathology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.
Methods Mol Biol. 2011;755:57-66. doi: 10.1007/978-1-61779-163-5_4.
Over the past 15 years, laser-based microdissection has improved the precision by which scientists can procure cells of interest from a heterogeneous tissue section. However, for studies that require a large amount of material (e.g., proteomics) or for cells that are scattered and difficult to identify by standard histological stains, an immunostain-based, automated approach becomes essential. In this chapter, we discuss the use of immunohistochemistry (IHC) and immunofluorescence (IF) to guide the microdissection process via manual and software-driven auto-dissection methods. Although technical challenges still exist with these innovative approaches, we present here methods and protocols to successfully perform immuno-based microdissection on commercially available laser dissection systems.
在过去的15年里，基于激光的显微切割技术提高了科学家从异质组织切片中获取感兴趣细胞的精度。然而，对于需要大量材料的研究（如蛋白质组学）或对于那些分散且难以通过标准组织学染色识别的细胞来说，基于免疫染色的自动化方法变得至关重要。在本章中，我们将讨论如何使用免疫组织化学（IHC）和免疫荧光（IF），通过手动和软件驱动的自动切割方法来指导显微切割过程。尽管这些创新方法仍然存在技术挑战，但我们在此介绍一些方法和方案，以便在商用激光切割系统上成功进行基于免疫的显微切割。
