It should be noted that all observations are derived from our experience with the beta version of the DSP instrument, which we are currently testing, and may not apply fully to the commercially available instrument. The working procedure of protein DSP DSP technology simultaneously characterizes regional and quantitative protein expression of up to 40 markers related to immune cell activation and tumor cell classification on a single FFPE tissue section . this article, we will describe our experience with the GeoMx DSP platform using cancer FFPE tissues. These expression profiles will provide better characterization and understanding of tumor heterogeneity and the tumor micro-environment, enabling the improvement of patient therapy and the identification of potential biomarker signatures. The L-779450 purpose of this article is to offer potential future users an independent insight into the L-779450 DSP platform and a comprehensive idea of usability, including advantages and current limitations of the technology based on our current experience with the beta version of NanoString’s DSP platform as part of the DSP beta-testing program. The GeoMxTM?Digital Spatial Profiling (DSP) platform is a non-destructive technique for regional in-depth protein expression profiling. Using oligonucleotide detection technologies, the GeoMxTM?DSP enables simultaneous high-level multiplexing on a single FFPE tissue. Here, we focus on our current experience derived from our biomarker research using the beta version of the DSP instrument. strong class=”kwd-title” Keywords: NanoString, DSP, Region of interest, Multiplex, IHC, Translational research Background Regional and quantitative protein signature analysis of clinical formalin-fixed paraffin-embedded (FFPE) tissues has proved to be difficult to perform in many research areas. In particular, in the field of immuno-oncology (IO), detailed characterization of immune cell subsets within a tumor area would improve our understanding of antitumor immunity and resistance to immune checkpoint inhibition. Tumors are highly heterogeneous in their architecture, (immune) cell composition, abundance and distribution. Understanding tumor and L-779450 immune cell co-localization may also be important for biomarker identification and precision immunotherapy , , , , . At present, the CD8+ T cell:regulatory T cell ratio is an improved biomarker compared with CD8 expression alone , , , , , . Furthermore, characterizing co-expression of co-stimulatory and inhibitory receptors would improve therapeutic strategies. For example, characterization of inhibitory markers expressed on T cells (e.g. LAG-3 and TIM-3) in the tumor micro-environment may impact the decision for new (combination) therapies , . Detailed tissue analysis is highly necessary but involves a laborious staining procedure and is hampered by limited patient tissue samples. Therefore, multiplex immunohistochemistry (IHC) will provide a more comprehensive insight into the interaction and crosstalk between tumor and immune cells within the tumor micro-environment. Multiplexing more than five antibodies has largely been restricted by the spectral overlap of available fluorophores or chromogens. Moreover, current multiplex IHC tools provide objective L-779450 information about the presence L-779450 and histological location of immune cells, but neglect detailed phenotypical description. Because of this limitation, additional circulation cytometry analysis is required. To advance standard IHC-based multiplex cells analyses, NanoString Systems (Seattle, WA) developed a novel imaging and tissue-sampling platform: GeoMxTM digital spatial profiling (DSP). This is a high-level multiplexing technique that provides information about the presence and histological location of immune cell subsets, and detailed characterization of their activation, differentiation and immune checkpoint manifestation . The capability of DSP technology in the field of Rabbit polyclonal to FASTK IO holds promise in advancing the current standard analysis of tumor biopsies. Therefore, this short article aims to give an insight into the DSP technology, the current recommended experimental set-up and an independent view on the advantages and limitations of the system based on our own encounter and study. It should be noted that all observations are derived from our encounter with the beta version of the DSP instrument, which we are currently testing, and may not apply fully to the commercially available instrument. The working process of protein DSP DSP technology simultaneously characterizes regional and quantitative protein expression of up to 40 markers related to immune cell activation and tumor cell classification on a single FFPE cells section . The DSP process implements five non-destructive steps: a standard FFPE tissue preparation step, a cells incubation step with.