Correlations Between Single Cell Signaling Dynamics and Protein Expressions Profiles

摘要

A platform technology for monitoring signaling pathways in single T cells using optical nanoprobes has been developed to provide a stable and biocompatible environment for the cells, and allow acquisition of additional data on cellular metabolic and physiological activity using other assays. Platform components were developed and validated to study the signaling dynamics of multiple T cells. MALDI-TOF protein detection, OCIBD IL2 quantification, and the use of quantum dots in live cell imaging were optimized for single cell applications. MALDI data showed clear activation dependence, but there was insufficient MALDI data for statistical analysis; ongoing technical developments are addressing this. Future efforts will fully integrate the microfluidic nanophysiometer, OCIBD analyze detection system, MALDI-TOF protein traps, and cell loading with quantum dots to achieve a fully-integrated, self-contained and instrumented cellular microenvironment that supports precise and timely decisions regarding when best to probe the cell invasively for more detailed intracellular signaling information. This system will allow the identification of unique patterns of signaling pathways of stimuli that can be used to identify previously unknown or maliciously engineered toxins, pathogens, antidotes, and prophylactics.