Enhanced Analysis of Tumor Microenvironment and Immune Regulation via an Automated Adjustable Signal Amplification Technique for Multiplex Immunofluorescence

Multiplex immunofluorescence (mIF) has become fundamental for tumor microenvironment (TME) and biomarker research [1]. Therein, different marker expression levels often have
biological meaning, and the detection of markers expressed at low vs. high levels can provide crucial insights [2]. However, detecting antigens expressed at lower levels can be challenging and often benefits from signal amplification [3]. Here, we demonstrate a new automated method for COMET™ instrument called sequential layered amplification (seqLA™) enabling the amplification of individual markers within multiplex panels. This technique provides an adjustable level of amplification and an efficient elution for subsequent staining cycles.