Feb 13, 2026
Version 1
Fabrication of alginic acid sodium salt -based MWC-chip V.1
- Zhe Zhao1
- 1CAS Key Laboratory of Nano-Bio Interface, Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences, Jiangsu, 215123, China.
- MTS
Protocol Citation: Zhe Zhao 2026. Fabrication of alginic acid sodium salt -based MWC-chip. protocols.io https://dx.doi.org/10.17504/protocols.io.dm6gp12w1gzp/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: February 13, 2026
Last Modified: February 13, 2026
Protocol Integer ID: 243161
Keywords: chip pdms micromolds for mts generation, chip pdms micromold, microfabrication of the mwc, fabrication of alginic acid sodium salt, microfabrication, alginic acid sodium salt, mts generation
Funders Acknowledgements:
National Natural Science Foundation of China
Grant ID: 82202345
Abstract
PDMS micromolds for MTS generation and microfabrication of the MWC-Chip.
Troubleshooting
The creation of PDMS male mold
The alginic acid sodium salt-based MWC-chips were fabricated using a micromolding method. Specifically, the layout of the chips was initially designed using AutoCAD software. A polymethyl methacrylate (PMMA) female mold was fabricated on a PMMA plate (CREROMEM, 200 mm × 300 mm × 2 mm) using a precision micromilling machine (JINGYAN) equipped with a parabolic spiral-flute bit (WeiTol, Φ0.5 mm). Subsequently, a polydimethylsiloxane (PDMS) male mold was created by pouring PDMS prepolymer (ratio 1:10, cross-linking agent: elastomer) into PMMA female mold and solidifying the PDMS prepolymer at 80 °C for 2 hours.
The generation of MWC-Chip
The released PDMS mold was then covered with an alginic acid sodium salt solution (2% w/v) (Sigma). The alginic acid sodium salt-based MWC-chips were formed by gelation in the CaCl2 solution (5% w/v) (Sinopharm Chemical Reagent) at 25 °C for 1 minutes and subsequently released from the PDMS mold. The MWC-chips were gently placed into a 3.5 cm2 plate. Each MWC-chip with a hydrogel substrate measured 2 × 2 cm2 and contained approximately 15,000 microwells with dimensions of diameter: 80 µm, height: 40 µm and pitch: 80 µm. The resulting MWC-chips were sterilized thoroughly in the purified PBS supplemented with penicillin-streptomycin at a concentration of 1% w/v, followed by exposure to ultraviolet light for over 30 minutes. Finally, these sterilized MWC-chips were stored at 4 °C for subsequent generation of micro-tumorspheres (MTSs).
Protocol references
Brandenberg, N. et al. High-throughput automated organoid culture via stem-cell aggregation in microcavity arrays. Nat Biomed Eng 4, 863-874 (2020).