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. 2019 Aug 9;10(8):527.
doi: 10.3390/mi10080527.

Microfluidic Generation of Amino-Functionalized Hydrogel Microbeads Capable of On-Bead Bioassay

Seongsoo Kim  1 , Sang-Myung Lee  1 , Sung Sik Lee  2   3 , Dong-Sik Shin  4
Affiliations

Microfluidic Generation of Amino-Functionalized Hydrogel Microbeads Capable of On-Bead Bioassay

Seongsoo Kim et al. Micromachines (Basel). .

Abstract

Microfluidic generation of hydrogel microbeads is a highly efficient and reproducible approach to create various functional hydrogel beads. Here, we report a method to prepare crosslinked amino-functionalized polyethylene glycol (PEG) microbeads using a microfluidic channel. The microbeads generated from a microfluidic device were evaluated by scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS) and confocal laser scanning microscopy, respectively. We found that the microbeads were monodisperse and the amino groups were localized on the shell region of the microbeads. A swelling test exhibited compatibility with various solvents. A cell binding assay was successfully performed with RGD peptide-coupled amino-functionalized hydrogel microbeads. This strategy will enable the large production of the various functional microbeads, which can be used for solid phase peptide synthesis and on-bead bioassays.

Keywords: hydrogel microbead; microfluidic channel; on-bead bioassay; peptide synthesis; polyethylene glycol.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Design of microfluidic channels for the generation of PEG-amine microbeads: (a) Non-functionalized PEG microbeads using two inlet channels and (b) amino-functionalized PEG-amine microbeads using three inlet channels. (b1) Flow focus junction, (b2) hydrogel precursor droplets flowing in channel, and (b3) emulsified droplets at the outlet.
Figure 2
Figure 2
Microscopic images of (a) emulsified hydrogel precursor droplets and (b) PEG-amine microbeads after destabilization. Scanning electron microscopic (SEM) images of (c) PEG-amine microbeads and (d) non-functionalized PEG microbeads. (e) Size distribution of PEG-amine microbeads (100 ea) and (f) microscopic image of PEG-amine microbeads swollen in water.
Figure 3
Figure 3
(a) Microscopic image of PEG-amine microbeads after bromophenol blue (BPB) staining. (b) Confocal laser scanning microscopic (CLSM) images of fluorescein isothiocyanate (FITC)-labeled PEG-amine microbeads at different z-levels and (c) three-dimensional reconstructed z-stack image.
Figure 4
Figure 4
Energy dispersive X-ray spectroscopy (EDS) analysis of PEG-amine microbeads (black) and non-functionalized PEG microbeads (red).
Figure 5
Figure 5
(a,d) Microscopic images and (b,c,e,f) confocal laser scanning microscope images of cell- treated hydrogel microbeads: NIH 3T3 cells were not attached on the PEG-amine beads (top) while cells were attached on GRGDSC peptide-immobilized PEG-amine microbeads (bottom).
See this image and copyright information in PMC

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