Ring-Shaped Micropatterning Cell Chirality Assay: An In Vitro Technique to Determine Multicellular Chirality Based on Cell Alignment on Ring-Shaped Micropatterns
Published: April 30, 2023
Abstract
Source: Zhang, H. et al. A Micropatterning Assay for Measuring Cell Chirality. J. Vis. Exp. (2022)
In this video, we describe a cell chirality assay to determine the alignment bias of cells in a confined geometric boundary such as a ring micropattern. Chirality is an inherent property of most cell types and is determined by genetic and environmental factors. Knowing the chirality of a normal cell population can help to screen drug-treated cells.
Protocol
1. Fabrication of polydimethylsiloxane (PDMS) stamps Draw an array of microscale rings using CAD software, with an inner diameter of 250 µm and an outer diameter of 450 µm. The pattern used in this protocol is a 10 x 10 array with an 850 µm distance between rings. Print a transparency mask of the pattern at the desired resolution using a microfabrication company's mask printing service (see Table of Materials). NOTE: The provided dimensions…
Representative Results
Figure 1. Schematic of cellular micropatterning. (A) Procedure of microfabrication and microcontact printing for cell patterning. A negative photoresist mold was made by ultraviolet (UV) crosslinking of photoresist via a mask containing micropatterning features (1-2). Polydimethylsiloxane (PDMS) elastomeric prepolymers were cast onto the mold to create stamps (3-4). Then, an adhe…
Offenlegungen
The authors have nothing to disclose.
Materials
| 200 proof ethanol | Koptec | DSP-MD-43 | |
| BZX microscope system | Keyence | BZX-600 | |
| Dulbecco's modified eagle medium (DMEM), high glucose | Gibco | 11965092 | |
| Electron beam evaporator | Temscal | BJD-1800 | Gold-titanum film coating |
| Fetal bovine serum | VWR | 89510-186 | |
| Fibronectin from bovine plasma | Sigma | F1141-5MG | |
| Glass microscope slides | VWR | 10024-048 | |
| Glass tweezers | Exelta | 390BSAPI | |
| Gold evaporation pellets | International Advanced Materials | AU18 | |
| HS-(CH2)11-EG3-OH (EG3) | Prochimia | TH 001-m11.n3-0.2 | |
| MATLAB | Mathworks | MATLAB_R2020b | |
| NIH/3T3 cells | ATCC | CRL-1658 | |
| OAI contact aligner | OAI | 200 | UV photolithography |
| Octadecanethiol (C18) | Sigma | O1858-25ML | |
| Orbital shaker | VWR | 89032-088 | |
| Phosphate buffered saline (PBS) | Research product international | P32080-100T | |
| Polydimethylsiloxane Sylgard 184 | Dow Corning | DC4019862 | |
| Silicon Wafer | University Wafer | ID#809 | |
| Sodium pyruvate | Thermo fisher scientific | 11360-070 | |
| SU-8 3050 photoresist | MicroChem | Y311075 0500L1GL | |
| Titanium evaporation pellets | International Advanced Materials | TI14 | |
| Transparency mask (with feature) | Outputicity.com | N/A | Mask printing service |
| Trypsin-EDTA (0.25%) | Thermo fisher scientific | 25200-072 |
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Ring-Shaped Micropatterning Cell Chirality Assay: An In Vitro Technique to Determine Multicellular Chirality Based on Cell Alignment on Ring-Shaped Micropatterns. J. Vis. Exp. (Pending Publication), e21153, doi: (2023).