Abstract
Single-cell proteomics (SCP) has advanced significantly in recent years,
with new tools that have been specifically designed for preparation and
analysis of single cells now commercially available to researchers. The
field is sufficiently mature to be broadly accessible to any lab capable
of isolating single cells and performing bulk-scale proteomic analyses.
In this review, we highlight recent work in the SCP field that has
significantly lowered the barrier to entry, thus providing a practical
guide for those who are newly entering the SCP field. We outline the
fundamental principles and report multiple paths to accomplish the key
steps of a successful SCP experiment including sample preparation,
separation and mass spectrometry data acquisition and analysis. We
recommend that researchers start with a label-free SCP workflow, as
achieving high-quality and quantitatively accurate results is more
straightforward than label-based multiplexed strategies. By leveraging
these accessible means, researchers can confidently perform SCP
experiments and make meaningful discoveries at the single-cell level.