We present a new and versatile implementation of rapid and localized immunohistochemical staining of tissue sections. Immunohistochemistry (IHC) comprises a sequence of specific biochemical reactions and allows the detection of specific proteins in tissue sections. For the rapid implementation of IHC, we fabricated horizontally oriented microfluidic probes (MFPs) with functionally designed apertures to enable square and circular footprints, which we employ to locally expose a tissue to time-optimized sequences of different biochemicals. We show that the two main incubation steps of IHC protocols can be performed on MDAMB468-1510A cell block sections in less than 30 min, compared to incubation times of an hour or more in standard protocols. IHC analysis on the timescale of tens of minutes could potentially be applied during surgery, enabling clinicians to react in more dynamically and efficiently. Furthermore, this rapid IHC implementation along with conservative tissue usage has strong potential for the implementation of multiplexed assays, allowing the exploration of optimal assay conditions with a small amount of tissue to ensure high-quality staining results for the remainder of the sample.

我们提出了一种新的、多功能的组织切片快速局部免疫组织化学染色方法。免疫组织化学 (IHC) 包括一系列特定的生化反应，可以检测组织切片中的特定蛋白质。为了快速实施 IHC，我们制造了具有功能设计孔径的水平取向微流体探针 (MFP)，以实现方形和圆形足迹，我们利用它们将组织局部暴露于不同生化物质的时间优化序列。我们表明，与标准协议中一小时或更长时间的孵育时间相比，IHC 协议的两个主要孵育步骤可以在不到 30 分钟的时间内在 MDAMB468-1510A 细胞块切片上执行。数十分钟的时间尺度上的 IHC 分析可能适用于手术期间，使临床医生能够更动态、更有效地做出反应。此外，这种快速的 IHC 实施以及保守的组织使用对于实施多重检测具有强大的潜力，允许使用少量组织探索最佳检测条件，以确保剩余样本的高质量染色结果。