The ability to visualize and quantify the spatial arrangement and geographic proximity of immune cells with tumor cells provides valuable insight into the complex mechanisms underlying cancer biology and progression. Multiplexing, which involves immunofluorescence labeling and the visualization of multiple epitopes within formalin-fixed paraffin embedded tissue sections, is a methodology that is being increasingly employed. Despite the power of immunofluorescence multiplex analysis, application of this technology to bone marrow core biopsies has not yet been realized. Given our specific long term goal to identify immune cells in proximity to bone marrow malignant plasma cells in multiple myeloma patients, we describe in this study adaptation of multiplex immunofluorescence analysis to this tissue. We first identified a blocking strategy that quenched autofluorescence. We next employed a multiplex strategy that uses a simple stripping solution to remove primary and secondary antibodies prior to subsequent rounds of staining. This method was found to be highly efficient and did not significantly alter antigenicity or tissue integrity. Our studies illustrate for the first time that immunofluorescence multiplexing is achievable in bone marrow core biopsies and will provide a novel opportunity to analyze the role of the immune contexture in disease progression of the monoclonal gammopathies.

中文翻译：

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骨髓核心活检的多重免疫荧光：可视化骨髓免疫情况。

可视化和量化免疫细胞与肿瘤细胞的空间排列和地理接近性的能力，为深入了解癌症生物学和进展的复杂机制提供了宝贵的见识。涉及免疫荧光标记和福尔马林固定石蜡包埋的组织切片内多个表位的可视化的多重化是一种越来越被采用的方法。尽管具有免疫荧光多重分析的功能，但尚未实现将该技术应用于骨髓核活检。鉴于我们确定多发性骨髓瘤患者骨髓恶性浆细胞附近的免疫细胞的特定长期目标，我们在本研究中描述了对该组织进行多重免疫荧光分析的适应性。我们首先确定了一种阻断自身荧光的阻断策略。接下来，我们采用了多重策略，该策略使用简单的剥离溶液在随后的几轮染色之前去除一抗和二抗。发现该方法是高效的，并且不会显着改变抗原性或组织完整性。我们的研究首次表明，在骨髓核心活检组织中可以实现免疫荧光多路复用，这将为分析免疫环境在单克隆配子体病发展中的作用提供新的机会。发现该方法是高效的，并且不会显着改变抗原性或组织完整性。我们的研究首次表明，在骨髓核心活检组织中可以实现免疫荧光多路复用，这将为分析免疫环境在单克隆同性恋病的疾病进展中的作用提供新的机会。发现该方法是高效的，并且不会显着改变抗原性或组织完整性。我们的研究首次表明，在骨髓核心活检组织中可以实现免疫荧光多路复用，这将为分析免疫环境在单克隆配子体病发展中的作用提供新的机会。