Creatv MicroTech, Inc., 1 Deer Park Dr. Monmouth Junction, NJ 08850, USA.
Fox Chase Cancer Center, Protocol Support Laboratory, 333 Cottman Ave., Philadelphia, PA 19111, USA.
Sci Rep. 2016 Sep 20;6:33488. doi: 10.1038/srep33488.
In tissue biopsies formalin fixed paraffin embedded cancer blocks are micro-sectioned producing multiple semi-identical specimens which are analyzed and subtyped proteomically, and genomically, with numerous biomarkers. In blood based biopsies (BBBs), blood is purified for circulating tumor cells (CTCs) and clinical utility is typically limited to cell enumeration, as only 2-3 positive fluorescent markers and 1 negative marker can be used. As such, increasing the number of subtyping biomarkers on each individual CTC could dramatically enhance the clinical utility of BBBs, allowing in depth interrogation of clinically relevant CTCs. We describe a simple and inexpensive method for quenching the specific fluors of fluorescently stained CTCs followed by sequential restaining with additional biomarkers. As proof of principle a CTC panel, immunosuppression panel and stem cell panel were used to sequentially subtype individual fluorescently stained patient CTCs, suggesting a simple and universal technique to analyze multiple clinically applicable immunomarkers from BBBs.
在组织活检中，福尔马林固定石蜡包埋的癌症块被微切片，产生多个半相同的标本，这些标本在蛋白质组学和基因组学上进行分析和亚型分析，并使用许多生物标志物。在基于血液的活检（BBB）中，血液被净化以用于循环肿瘤细胞（CTC），并且临床应用通常仅限于细胞计数，因为仅可以使用 2-3 个阳性荧光标记物和 1 个阴性标记物。因此，增加每个个体 CTC 的亚型生物标志物数量可以显著提高 BBB 的临床实用性，从而深入研究临床相关的 CTC。我们描述了一种简单且廉价的方法，用于淬灭荧光染色的 CTC 的特定荧光，然后用其他生物标志物进行顺序重新染色。作为原理验证，使用 CTC 面板、免疫抑制面板和干细胞面板对单个荧光染色的患者 CTC 进行顺序亚型分析，这表明一种简单而通用的技术可用于分析来自 BBB 的多个临床适用的免疫标志物。
