Ring-Shaped Micropatterning Cell Chirality Assay: An In Vitro Technique to Determine Multicellular Chirality Based on Cell Alignment on Ring-Shaped Micropatterns

In This Article

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 of the ring have been proven to work for many cell types.
Conduct ultraviolet (UV) photolithography using the mask containing desired features ....

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Results

Photolithography process diagram and experimental steps for PDMS stamp fabrication and cell seeding.
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.......