Microfluidic based single cell or droplet manipulation: Methods and applications.

The isolation of single cell or droplet is first and crucial step to single-cell analysis, which is important for cancer research and diagnostic methods. This review provides an overview of technologies that are currently used or in development to realize the isolation. Microfluidic based manipulation is an emerging technology with the distinct advantages of miniaturization and low cost. Therefore, recent developments in microfluidic isolated methods have attracted extensive attention. We introduced herein five strategies based on microfluid: trap, microfluidic discrete manipulation, bioprinter, capillary and inertial force. For every technology, their basic principles and features were discussed firstly. Then some modified approaches and applications were listed as the extension. Finally, we compared the advantages and drawbacks of these methods, and analyzed the trend of the manipulation based on microfluidics.

Droplet magnetic-controlled microfluidic chip integrated nucleic acid extraction and amplification for the detection of pathogens and tumor mutation sites.

Traditional nucleic acid extraction and detection is based on open operation, which may cause cross-contamination and aerosol formation. This study developed a droplet magnetic-controlled microfluidic chip integrated nucleic acid extraction, purification and amplification. The reagent is sealed in oil to form a droplet, and the nucleic acid is extracted and purified by controlling the movement of the magnetic beads (MBs) through a permanent magnet, ensuring a closed environment. This chip can automatically extract nucleic acid from multiple samples within 20 min, and can be directly placed in the in situ amplification instrument for amplification without further transfer of nucleic acid, characterized by simple, fast, time-saving and labor-saving. The results showed that the chip was able to detect <10 copies/test SARS-CoV-2 RNA, and EGFR exon 21 L858R mutations were detected in H1975 cells as low as 4 cells. In addition, on the basis of the droplet magnetic-controlled microfluidic chip, we further developed a multi-target detection chip, which used MBs to divide the nucleic acid of the sample into three parts. And the macrolides resistance mutations A2063G and A2064G, and the P1 gene of mycoplasma pneumoniae (MP) were successfully detected in clinical samples by the multi-target detection chip, providing the possibility for future application in the detection of multiple pathogens.

The Discovery of Novel Circulating Cancer-Related Cells in Circulation Poses New Challenges to Microfluidic Devices for Enrichment and Detection.

Circulating tumor cells (CTCs) enumeration has been widely used as a surrogate predictive marker for early diagnoses, the evaluation of chemotherapy efficacy, and cancer prognosis. Microfluidic technologies for CTCs enrichment and detection have been developed and commercialized as automation platforms. Currently, in addition to CTCs, some new types of circulating cancer-related cells (e.g., CCSCs, CTECs, CAMLs, and heterotypic CTC clusters) in circulation are also reported to be correlated to cancer diagnosis, metastasis, or prognosis. And they widely differ from the conventional CTCs in positive markers, cellular morphology, or size, which presents a new technological challenge to microfluidic devices that use affinity-based capture methods or size-based filtration methods for CTCs detection. This review focuses on the biological and physical properties as well as clinical significance of the novel circulating cancer-related cells, and discusses the challenges of their discovery to microfluidic chip for enrichment. Finally, the current challenges of CTCs detection in clinical application and future opportunities are also discussed.

Synthesis of carbon quantum dots with iron and nitrogen from Passiflora edulis and their peroxidase-mimicking activity for colorimetric determination of uric acid.

Carbon quantum dots co-doped with iron and nitrogen (Fe@NCDs) were synthesized by using Passiflora edulis Sims (P. edulis) as a precursor. The Fe@NCDs exhibit outstanding peroxidase-mimetic activity owing to successful doping with iron resulting in a behavior similar to that of iron porphyrins. In the presence of H2O2, the Fe@NCDs catalyze the oxidation of the peroxidase substrate 3,3',5,5'-tetramethylbenzidine (TMB) with a color change from colorless to blue. The blue oxidation product has a characteristic absorption peaking at 652 nm. A colorimetric assay was worked out for uric acid (UA) that measures the hydrogen peroxide produced during oxidation of UA by uricase. Response is linear in the 2-150 µM UA concentration range, and the limit of detection is 0.64 µM. The method was applied to the determination of UA in (spiked) urine, and recoveries ranged from 92.0 to 103.4%. Graphical abstract Schematic representation of the fabrication of iron and nitrogen co-doped carbon dots (Fe@NCDs) using Passiflora edulis Sims as carbon-based materials. First, uric acid (UA) was oxidized to generate H2O2 by uricase. Then, the Fe@NCDs catalyzed the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) to form blue-colored oxidized TMB (oxTMB) in the presence of H2O2. UA can be quantified based on the theory.