Abstract by Samuel Squires
Chemistry and Biochemistry
Optimization of Sample Preparation Technique for Single Cell Proteomics
Mass spectrometry based proteomics has become a powerful approach for resolving cellular heterogeneity, which is crucial for understanding cellular and disease mechanisms. The recently developed Nanodroplet Processing in One pot for Trace Samples (nanoPOTS) workflow enables high-resolution proteome coverage of small-cell populations. Despite its application in a number of nanoscale proteomic studies, however, the fundamentals of the technique have remained unchanged for single-cell sample processing. Here, we aim to optimize sample processing to achieve a depth of coverage of >3000 proteins/cell for single-cell studies. By manipulating sample volume, nanowell surface area, and reagent concentrations, we will achieve shorter processing times and increased sample recovery, enabling high-throughput proteomic analysis of single cells with far greater efficiency. As a result, higher resolution of cellular heterogeneity can be achieved in less time, providing an enhanced tool for studying, diagnosing, and guiding treatments for disease.