Kim Minseok S, Kwon Seyong, Park Je-Kyun
Department of Bio and Brain Engineering, Korea Advanced Institute of Science and Technology (KAIST), Yuseong-gu, Daejeon, Republic of Korea.
Methods Mol Biol. 2013;949:349-64. doi: 10.1007/978-1-62703-134-9_22.
A quantitative, reproducible, fast and inexpensive multiplexed immunohistochemistry (IHC) system might play a locomotive role in drug screening and personalized medicine. Currently, fully automated IHC machines and sequential multiplexed IHC methods based upon multiple color reagents have been developed, with the evolution of such methods having revealed novel biological findings over the conventional IHC method, which is time consuming and labor intensive. We describe a novel parallel multiplexed IHC method using a microfluidic multiplexed immunohistochemistry (MMIHC) device for quantitative pathological diagnosis of breast cancer. The key factors for success of parallel multiplexed IHC are the fabrication of a robust microfluidic device, the interface between the device and a tissue slide, and an accurate fluidic control for multiple IHC reagents. In order to apply conventional thin-section tissues into on-chip systems without any additional modification process, a tissue slide-compatible assembler was developed for optimal compatibility of conventional IHC methods. With this approach, a perfect fluid control for various solutions was demonstrated without any leakage, bubble formation or cross-contamination. The results presented in this chapter indicate that the microfluidic IHC protocol developed can provide the possibility of tailored cancer treatments as well as precise histopathological diagnosis using numerous specific biomarkers.
一种定量、可重复、快速且廉价的多重免疫组织化学（IHC）系统可能在药物筛选和个性化医疗中发挥引领作用。目前，随着基于多种颜色试剂的全自动免疫组化机器和顺序多重免疫组化方法的发展，这些方法的演变揭示了一些比传统免疫组化方法更新颖的生物学发现，传统免疫组化方法既耗时又费力。我们描述了一种使用微流控多重免疫组化（MMIHC）装置进行乳腺癌定量病理诊断的新型平行多重免疫组化方法。平行多重免疫组化成功的关键因素包括坚固的微流控装置的制造、装置与组织切片之间的界面以及对多种免疫组化试剂的精确流体控制。为了在无需任何额外修饰过程的情况下将传统薄切片组织应用于芯片系统，开发了一种与组织切片兼容的组装器，以实现传统免疫组化方法的最佳兼容性。通过这种方法，展示了对各种溶液的完美流体控制，没有任何泄漏、气泡形成或交叉污染。本章给出的结果表明，所开发的微流控免疫组化方案能够利用众多特定生物标志物实现定制化癌症治疗以及精确的组织病理学诊断。
