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. 2023 May;17(3):197-203.
doi: 10.1049/nbt2.12114. Epub 2023 Jan 16.

Optically driven microtools with an antibody-immobilised surface for on-site cell assembly

Shuntaro Mori  1 , Takumi Ito  1 , Hidekuni Takao  1   2 , Fusao Shimokawa  1   2 , Kyohei Terao  1   2
Affiliations

Optically driven microtools with an antibody-immobilised surface for on-site cell assembly

Shuntaro Mori et al. IET Nanobiotechnol. 2023 May.

Abstract

To enable the accurate reproduction of organs in vitro, and improve drug screening efficiency and regenerative medicine research, it is necessary to assemble cells with single-cell resolution to form cell clusters. However, a method to assemble such forms has not been developed. In this study, a platform for on-site cell assembly at the single-cell level using optically driven microtools in a microfluidic device is developed. The microtool was fabricated by SU-8 photolithography, and antibodies were immobilised on its surface. The cells were captured by the microtool through the bindings between the antibodies on the microtool and the antigens on the cell membrane. Transmembrane proteins, CD51/61 and CD44 that facilitate cell adhesion, commonly found on the surface of cancer cells were targeted. The microtool containing antibodies for CD51/61 and CD44 proteins was manipulated using optical tweezers to capture HeLa cells placed on a microfluidic device. A comparison of the adhesion rates of different surface treatments showed the superiority of the antibody-immobilised microtool. The assembly of multiple cells into a cluster by repeating the cell capture process is further demonstrated. The geometry and surface function of the microtool can be modified according to the cell assembly requirements. The platform can be used in regenerative medicine and drug screening to produce cell clusters that closely resemble tissues and organs in vivo.

Keywords: bioMEMS; laser beam applications; microchannel flow; microfabrication.

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Conflict of interest statement

There are no conflicts to declare.

Figures

FIGURE 1
FIGURE 1
Cell manipulation and assembly using an optically driven microtool with an antibody‐immobilised surface.
FIGURE 2
FIGURE 2
Fabrication process of microtools. The bottom scheme depicts the mechanism of immobilisation of antibodies on the SU‐8 surface.
FIGURE 3
FIGURE 3
Various shapes of Microtools. Upper: Design. Lower: SEM images. (a) Square, (b) circle, (c) rectangle, and (d) cross‐shaped microtools.
FIGURE 4
FIGURE 4
Microchannel for cell‐trapping and assembly. (a) Dimensions of the channel. (b) Introducing microtools into the storage chambers by absorption of air into polydimethylsiloxane (PDMS). (c) Introducing cells and single‐cell trapping. (d) Manipulating microtools by optical tweezers and cell assembly. (e) Photo of the microfluidic device.
FIGURE 5
FIGURE 5
Experimental setup for cell assembly using optical tweezers.
FIGURE 6
FIGURE 6
Cell manipulation with a microtool. Left: An optically driven microtool with CD51/61 antibodies in contact with a trapped cell. Right: Translocation of the cell through the microtool.
FIGURE 7
FIGURE 7
Comparison of adhesion efficiency between two surface treatments on microtools. The numbers in the bars show the number of the trials in the form of (successful adhesion)/(total trial).
FIGURE 8
FIGURE 8
Comparison of adhesion efficiency between 2 cell types. The numbers in the bars show the number of the trials in the form of (successful adhesion)/(total trial).
FIGURE 9
FIGURE 9
Cell assembly with microtools. (a) 3‐cell assembly with a square microtool. (b) 4‐cell assembly with a cross‐shape section microtool.
FIGURE 10
FIGURE 10
Cell assembly with microtools. (a) Assembly of a HeLa‐H2B‐GFP (left) and a MDA‐MB‐231 cell (right). The HeLa‐H2B‐GFP cell shows fluorescence in the cell nucleus. (b) 4‐cell straight assembly of Hela cells with 3‐microtools.
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