Second-Strand Synthesis-Based Massively Parallel scRNA-Seq Reveals Cellular States and Molecular Features of Human Inflammatory Skin Pathologies.

High-throughput single-cell RNA-sequencing (scRNA-seq) methodologies enable characterization of complex biological samples by increasing the number of cells that can be profiled contemporaneously. Nevertheless, these approaches recover less information per cell than low-throughput strategies. To accurately report the expression of key phenotypic features of cells, scRNA-seq platforms are needed that are both high fidelity and high throughput. To address this need, we created Seq-Well S3 ("Second-Strand Synthesis"), a massively parallel scRNA-seq protocol that uses a randomly primed second-strand synthesis to recover complementary DNA (cDNA) molecules that were successfully reverse transcribed but to which a second oligonucleotide handle, necessary for subsequent whole transcriptome amplification, was not appended due to inefficient template switching. Seq-Well S3 increased the efficiency of transcript capture and gene detection compared with that of previous iterations by up to 10- and 5-fold, respectively. We used Seq-Well S3 to chart the transcriptional landscape of five human inflammatory skin diseases, thus providing a resource for the further study of human skin inflammation.

Automated transportation of single cells using robot-tweezer manipulation system.

Manipulation of biological cells becomes increasingly important in biomedical engineering to address challenge issues in cell-cell interaction, drug discovery, and tissue engineering. Significant demand for both accuracy and productivity in cell manipulation highlights the need for automated cell transportation with integrated robotics and micro/nano manipulation technologies. Optical tweezers, which use highly focused low-power laser beams to trap and manipulate particles at micro/nanoscale, have emerged as an essential tool for manipulating single cells. In this article, we propose to use a robot-tweezer manipulation system to solve the problem of automatic transportation of biological cells, where optical tweezers function as special robot end effectors. Dynamics equation of the cell in optical tweezers is analyzed. A closed-loop controller is designed for transporting and positioning cells. Experiments are performed on live cells to demonstrate the effectiveness of the proposed approach in effective cell positioning.