Abstract: The present invention recognizes that current clinical laboratory testing methods for multiparametric single cell analysis are limited to analysis of intact live cells, and are insufficient for identification of signaling activation profile defining certain cell types, including but not limited to neoplastic and immunologically activated cell subsets. One aspect of the present invention generally relates to marker selection in panels to include proteins routinely assessed in standard FCM, while preferably also incorporating markers for surface receptor proteins within activated signaling cascades.

Abstract: Methods for preparation of cells for analysis of biomarkers are disclosed. In one aspect, the method for preparation of cells for analysis of biomarkers includes contacting a sample that contains a population of cells with at least one modulating substance at a first temperature, thereby producing a modulated cell population; contacting the modulated cell population with at least one antibody that is directed to a cell surface biomarker at a second temperature that is lower than the first temperature, thereby producing an extracellularly stained cell population; and contacting the extracellularly stained cell population with one or more reagents that fixes and permeabilizes the cells, thereby producing a fixed and permeabilized cell population.

Abstract: The present invention recognizes that current clinical laboratory testing methods for multiparametric single cell analysis are limited to analysis of intact live cells, and are insufficient for identification of signaling activation profile defining certain cell types, including but not limited to neoplastic and immunologically activated cell subsets. One aspect of the present invention generally relates to marker selection in panels to include proteins routinely assessed in standard FCM, while preferably also incorporating markers for surface receptor proteins within activated signaling cascades.

Abstract: Methods for preparation of cells for analysis of biomarkers are disclosed. In one aspect, the method for preparation of cells for analysis of biomarkers includes contacting a sample that contains a population of cells with at least one modulating substance at a first temperature, thereby producing a modulated cell population; contacting the modulated cell population with at least one antibody that is directed to a cell surface biomarker at a second temperature that is lower than the first temperature, thereby producing an extracellularly stained cell population; and contacting the extracellularly stained cell population with one or more reagents that fixes and permeabilizes the cells, thereby producing a fixed and permeabilized cell population.