Clickable Multifunctional Dumbbell Particles for in Situ Multiplex Single-Cell Cytokine Detection

Abstract

We report a novel strategy for fabrication of multifunctional dumbbell particles (DPs) through click chemistry for monitoring single-cell cytokine releasing. Two different types of DPs were prepared on a large scale through covalent bioorthogonal reaction between methyltetrazine and trans-cyclooctene on a microchip under a magnetic field. After collection of the DPs, the two sides of each particle were further functionalized with different antibodies for cell capturing and cytokine detection, respectively. These DPs labeled with different fluorescent dyes have been used for multiplex detection and analysis of cytokines secreted by single live cells. Our results show that this new type of DPs are promising for applications in cell sorting, bioimaging, single-cell analysis, and biomedical diagnostics.

Highly Multiplexed Single‐Cell Protein Profiling with Large‐Scale Convertible DNA‐Antibody Barcoded Arrays

“…Clamping of the microchip prevents any leakage of cytokines between microwells during the 6 h incubation time (Figure S4, Supporting Information). A sufficient number of cytokines would be produced within 6 h according to our previous study and literature 19, 20. Longer incubation time may affect cell viability and stress cells to alter their functions, due to limited available nutrition in an enclosed microchamber.…”

“…Preparation of cDNA‐Antibody Conjugates : The conjugation of cDNA with antibody was described before 19. In brief, the 200 × 10 −6 m complementary oligonucleotides were modified with 4‐formylbenzamide group (SFB; Solulink), while the 3.3 × 10 −6 m antibodies were modified with 6‐hydrazinonicotinamide (s‐HyNic; Solulink).…”

Metallic Nanoparticle-Based Optical Cell Chip for Nondestructive Monitoring of Intra/Extracellular Signals

“…Recently, there have been a lot of cell chip platforms to allow in situ analysis of the several biomaterials such as DNA, miRNA, mRNA, cytokines, etc., in live cells due to their versatility for application to toxicological tests, drug screening, and investigation of stem cell fate [89][90][91][92][93][94]. Diverse detection methods have been integrated into the cell chip; however, most of the analytical methods require labeling of the signaling molecules such as fluorophore for the immunofluorescence or other fluorescence-based methods.…”

Magnetic Janus Particles for Static and Dynamic (Bio)Sensing

“…Regarding protein detection, multifunctional dumbbell-shaped magnetic Janus particles were prepared and modified on one face with anti-CD14 antibody for cell attachment and coated with anti-TNFα antibody on the other face ( Figure 7a). The system allowed specific cell targeting and in situ multi-detection of cytokines [46]. allow easy sample separation/concentration, and can be reused by simple isolation through magnetic separation [41].…”

“…Both sensing assays were integrated into the same Janus type particle, thus, allowing simultaneous monitoring of glucose and cholesterol in human serum. Janus magnetic nanoparticles have also been used for DNA [43,44] and protein detection [45,46]. Doyle's group brilliantly developed polymeric magnetic Janus particles with separate detection and barcoding compartments [44,47].…”