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Single Cell Isolation Using Optical Tweezers
Anusha Keloth1, Owen Anderson2, Donald Risbridger3
1Institute of Biological Chemistry, Biophysics and Bioengineering, School of Engineering and Physical Sciences, Heriot Watt University, Edinburgh EH14 4AS, UK. anushakeloth89@gmail.com.
Micromachines
|November 15, 2018
Summary
Optical tweezers enable precise single-cell manipulation for applications in biotechnology. This study demonstrates successful isolation and co-culture of yeast and bacteria, impacting industrial processes and pathogen dynamics research.
Area of Science:
- Biotechnology
- Microfluidics
- Cell Biology
Background:
- Optical tweezers provide a non-contact method for precise cell manipulation.
- Understanding single-cell behavior in defined microenvironments is crucial for various biological applications.
Purpose of the Study:
- To characterize optical tweezing of yeast (Saccharomyces cerevisiae) and evaluate different cell isolation devices.
- To assess the viability and budding behavior of yeast cells after optical tweezing.
- To construct micro-consortia and co-cultures for potential biotechnological and pathogen dynamics research.
Main Methods:
- Characterization of optical tweezing parameters (laser power, speed) for yeast cell manipulation.
- Fabrication and testing of three cell isolation devices: micropipette, PDMS chip, and laser-machined fused silica chip.
- Culturing and observation of isolated single yeast cells in PDMS chips for up to 18 hours.
- Construction of yeast micro-consortia and yeast-bacteria co-cultures using optical tweezers and PDMS devices.
Main Results:
- Single yeast cells were successfully manipulated at 0.41 ± 0.06 mm/s using a 785 nm diode laser (26.8 ± 0.1 mW).
- PDMS chips demonstrated the most effective cell isolation, allowing yeast growth for 18 hours without contamination.
- Yeast cell viability was confirmed after optical tweezing with specific laser parameters (25.0 ± 0.1 mW for 1 min).
- Increased laser energy prolonged the time to the first budding event in S. cerevisiae, but did not affect budding duration.
Conclusions:
- Optical tweezers, combined with PDMS microfluidic devices, offer a robust method for isolating and culturing single cells, including bacteria and cyanobacteria.
- The technology facilitates the creation of defined microbial communities, with implications for industrial biotechnology and the study of pathogen dynamics.
- Cell viability and biological processes like budding are maintained under optimized optical tweezing conditions.
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