Cilo-seq: highly sensitive cell-in-library-out single-cell transcriptome sequencing with digital microfluidics

Abstract

Single-cell RNA sequencing (scRNA-seq) plays a critical role in revealing genetic expression patterns at the single-cell level for cell type identification and rare transcript detection. Although there have been great...

Digital microfluidics for biological analysis and applications

“…71 Comparatively, this system enabled fewer than 10 copies of template to be detected with a hundred-fold improvement in Copyright 2022, The Royal Society of Chemistry. 69 the detection limit compared to a bench-top assay. Unlike LAMP and RPA, which could only amplify known DNA sequences, MDA could carry out whole genome amplification using a random primer and phi29 DNA polymerase.…”

“…8D). 69 After cDNA amplification, purification beads were utilized on chip to adsorb long-chain cDNAs, while leaving short-chain primers in the supernatant, facilitating the separation of cDNA products. Afterward, library preparation reagents were introduced in sequence for DNA fragmentation, adaptor addition, library amplification and magnet-aided product purification.…”

Microfluidic single‐cell multiomics analysis

“…With an array of electrodes, the picoliter‐to‐microliter droplets can be dispensed, moved, merged, and split programmatically according to the preset path. With the benefits of automation, flexibility, dynamic centrifugation capacity and low reagent consumption, DMF has been widely employed in DNA detection, 31 protein pretreatment, 32,33 and cell analysis 34 …”

“…With the benefits of automation, flexibility, dynamic centrifugation capacity and low reagent consumption, DMF has been widely employed in DNA detection, 31 protein pretreatment, 32,33 and cell analysis. 34 The hydrophilic sites on the DMF device are typically utilized for cell isolation and adhesion. 35,36 The Wheel group devised Digital microfluidic Isolation of Single Cells for Omics (DISCO) system for target single-cell lysis and cellular contents recovery (Figure 3A).…”

Optimization of Electrode Patterns for an ITO-Based Digital Microfluidic through the Finite Element Simulation

“…Based on the effect of electrowetting-on-dielectric (EWOD), the DMF is able to control the wetting behavior of droplets through an array of electrodes and manipulate the droplets to split, dispense, mix and transfer without mechanical accessories [ 2 , 3 , 4 ]. Recent advances in manufacturing and electronic techniques have significantly promoted the development of DMF systems to achieve advantages such as a low power consumption, low reagent cost, miniaturization, flexibility and reconfiguration [ 5 , 6 ] Hence, the applications of DMF are increasingly being extended to fields such as point-of-care testing [ 7 , 8 ], cell research [ 9 , 10 , 11 , 12 ], biomedicine [ 13 ] and environmental monitoring [ 14 , 15 ].…”