Department of Bio and Brain Engineering, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea.
KAIST Institute for Health Science and Technology, 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea.
Lab Chip. 2021 Sep 14;21(18):3471-3482. doi: 10.1039/d1lc00375e.
We present a multiplexed microfluidic immunohistochemistry (IHC) technology that enables high-throughput analysis of tissue microarrays (TMAs) using the patterns of biomarker barcodes, which consist of a series of expressed linear patterns of specific biomarkers. A multichannel poly(dimethylsiloxane) microfluidic device was reversibly assembled by the pressure of simple equipment for multiplexed IHC on each core of TMA or cell microarray (CMA) section slides. By injecting primary antibodies from different biomarkers independently into each channel, multiplexed immunostaining can be performed on each core of TMA. We confirmed the equal immunostaining quality regardless of the channel orders and core positions in the slide. Four different biomarkers (ER, PR, HER2, and Ki67) were used for the demonstration of distinctive expression patterns on CMAs which consist of six different breast cancer cell lines, and it was confirmed that these bar-like signals could be a biomarker barcode for the TMA core. A biomarker barcode of breast cancer patient-derived TMA was quickly scanned by a slide scanner and compared to the conventional method for breast cancer diagnosis. This "barcode-IHC" concept, which has been verified by performing multiplexed microfluidic IHC on CMA and TMA samples, provides high reproducibility and the potential of high-throughput screening with molecular diagnostic capability.
我们提出了一种多重微流控免疫组织化学（IHC）技术，该技术使用生物标志物条码模式对组织微阵列（TMA）进行高通量分析，这些条码模式由一系列特定生物标志物的表达线性模式组成。通过使用简单设备的压力，将多通道聚二甲基硅氧烷（PDMS）微流控装置可逆地组装到 TMA 或细胞微阵列（CMA）切片的每个核心上，用于进行多重 IHC。通过将来自不同生物标志物的一抗分别注入每个通道中，可以对 TMA 的每个核心进行多重免疫染色。我们证实了无论通道顺序和载玻片上的核心位置如何，免疫染色的质量都是相等的。使用四种不同的生物标志物（ER、PR、HER2 和 Ki67）对由六种不同乳腺癌细胞系组成的 CMA 进行了独特表达模式的演示，证实了这些条状信号可以作为 TMA 核心的生物标志物条码。通过对 CMA 和 TMA 样本进行多重微流控 IHC，可以快速扫描乳腺癌患者衍生的 TMA 的生物标志物条码，并与传统的乳腺癌诊断方法进行比较。这种已经通过在 CMA 和 TMA 样本上进行多重微流控 IHC 得到验证的“条码-IHC”概念，提供了高重复性和高通量筛选的潜力，具有分子诊断能力。
