Author:
Hyman Leland B.,Christopher Clare R.,Romero Philip A.
Abstract
AbstractExperimental methods that capture the individual properties of single cells are revealing the key role of cell-to-cell variability in countless biological processes. These single-cell methods are becoming increasingly important across the life sciences in fields such as immunology, regenerative medicine, and cancer biology. Existing single-cell analysis methods are often limited by their low analysis throughput, their inability to profile high-dimensional phenotypes, and complicated experimental workflows with slow turnaround times. In this work, we present Single-cell Nucleic Acid Profiling in Droplets (SNAPD) to analyze the transcriptional states of hundreds of thousands of single mammalian cells. Individual cells are encapsulated in aqueous droplets on a microfluidic chip and the content of each cell is profiled by amplifying a targeted panel of transcriptional markers. Molecular logic circuits then integrate this multi-dimensional information to categorize cells based on their transcriptional profile and produce a detectable fluorescence output. SNAPD analyzes over 100,000 cells per hour and can be used to quantify distinct cell types within populations, detect rare cells at frequencies down to 0.1%, and enrich specific cell types using microfluidic sorting. SNAPD provides a simple, rapid, low cost, and scalable approach to study complex phenotypes in heterogeneous cell populations.
Publisher
Cold Spring Harbor Laboratory