Department of Pathology and Laboratory Medicine, Weill Cornell Medicine, New York, NY, USA.
Commun Biol. 2022 Jun 27;5(1):627. doi: 10.1038/s42003-022-03558-8.
Multiparametric imaging allows researchers to measure the expression of many biomarkers simultaneously, allowing detailed characterization of cell microenvironments. One such technique, CODEX, allows fluorescence imaging of >30 proteins in a single tissue section. In the commercial CODEX system, primary antibodies are conjugated to DNA barcodes. This modification can result in antibody dysfunction, and development of a custom antibody panel can be very costly and time consuming as trial and error of modified antibodies proceeds. To address these challenges, we developed novel tyramide-conjugated DNA barcodes that can be used with primary antibodies via peroxidase-conjugated secondary antibodies. This approach results in signal amplification and imaging without the need to conjugate primary antibodies. When combined with commercially available barcode-conjugated primary antibodies, we can very quickly develop working antibody panels. We also present methods to perform antibody staining using a commercially available automated tissue stainer and in situ hybridization imaging on a CODEX platform. Future work will include application of the combined tyramide-based and regular CODEX approach to image specific tumors with their immune cell infiltrates, including biomarkers that are currently difficult to image by regular CODEX.
多参数成像是一种能够同时测量多种生物标志物表达水平的技术，它可以对细胞微环境进行详细的描述。其中一种技术 CODEX 可以在单个组织切片上进行 >30 种蛋白质的荧光成像。在商业 CODEX 系统中，一抗与 DNA 条码连接。这种修饰可能导致抗体功能失调，并且由于需要进行反复试验以修改抗体，因此开发定制的抗体面板可能非常昂贵且耗时。为了解决这些挑战，我们开发了新型的酪胺偶联 DNA 条码，它可以通过过氧化物酶偶联的二抗与一抗一起使用。这种方法无需连接一抗即可实现信号放大和成像。当与市售的带有条码的一抗结合使用时，我们可以非常快速地开发工作抗体面板。我们还介绍了使用市售的自动化组织染色机进行抗体染色以及在 CODEX 平台上进行原位杂交成像的方法。未来的工作将包括应用基于酪胺的联合和常规 CODEX 方法来对具有免疫细胞浸润的特定肿瘤进行成像，包括目前通过常规 CODEX 难以成像的生物标志物。
