Related Experiment Video
Updated: Jun 10, 2026
A Microfluidic Platform for High-throughput Single-cell Isolation and Culture
Published on: June 16, 2016
Microtable arrays for culture and isolation of cell colonies
Jeng-Hao Pai1, Wei Xu, Christopher E Sims
1Department of Chemistry, University of North Carolina, Chapel Hill, NC 27599, USA.
Abstract:
Cell microarrays with culture sites composed of individually removable microstructures or micropallets have proven benefits for isolation of cells from a mixed population. The laser energy required to selectively remove these micropallets with attached cells from the array depends on the microstructure surface area in contact with the substrate. Laser energies sufficient to release micropallets greater than 100 μm resulted in loss of cell viability. A new three-dimensional culture site similar in appearance to a table was designed and fabricated using a simple process that relied on a differential sensitivity of two photoresists to UV-mediated photopolymerization. With this design, the larger culture area rests on four small supports to minimize the surface area in contact with the substrate. Microtables up to 250 × 250 μm were consistently released with single 10-μJ pulses to each of the four support structures. In contrast, microstructures with a 150 × 150-μm surface area in contact with the substrate could not be reliably released at pulse energies up to 212 μJ. Cassie-Baxter wetting is required to provide a barrier of air to localize and sequester cells to the culture sites. A second asset of the design was an increased retention of this air barrier under conditions of decreased surface tension and after prolonged culture of cells. The improved air retention was due to the hydrophobic cavity created beneath the table and above the substrate which entrapped air when an aqueous solution was added to the array. The microtables proved an efficient method for isolating colonies from the array with 100% of selected colonies competent to expand following release from the array.
Insights
Researchers developed novel microtable cell culture sites for efficient cell isolation. This new design minimizes laser energy for release, preserving cell viability and enabling colony expansion.
Area of Science:
- Biotechnology and Biomedical Engineering
- Cell Biology and Tissue Engineering
Background:
- Cell microarrays with removable microstructures aid in isolating cells from mixed populations.
- Traditional methods using laser-based release of microstructures are limited by cell viability issues due to high energy requirements for larger surface areas.
Purpose of the Study:
- To design and fabricate a novel three-dimensional (3D) culture site for improved cell isolation from microarrays.
- To minimize laser energy required for microstructure release, thereby preserving cell viability.
- To enhance the retention of the air barrier for better cell sequestration and culture conditions.
Main Methods:
- Fabrication of a 3D microtable culture site using differential photoresist sensitivity to UV photopolymerization.
- The microtable design features a large culture area supported by small structures to reduce substrate contact.
- Evaluation of laser-induced release efficiency and cell viability at varying laser energies and microstructure sizes.
- Assessment of air barrier retention under different surface tension conditions and prolonged cell culture.
Main Results:
- Microtables up to 250 × 250 μm were reliably released using low-energy (10 μJ) laser pulses on support structures.
- Conventional microstructures with larger contact areas (150 × 150 μm) required significantly higher energies (up to 212 μJ) for release and showed cell viability loss.
- The microtable design demonstrated superior air barrier retention due to a hydrophobic cavity, even under reduced surface tension and extended culture periods.
- 100% of isolated colonies were competent for expansion after release from the microtable array.
Conclusions:
- The novel 3D microtable culture site offers an efficient and viable method for isolating cell colonies from microarrays.
- This design overcomes the limitations of previous methods by enabling low-energy release and maintaining cell integrity.
- The enhanced air barrier properties contribute to improved cell localization and culture stability.
Related Concept Videos
01:24Techniques for Isolation of Pure Cultures
01:20Overview Of Cell Separation And Isolation
01:24iChip