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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.

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Summary
This summary is machine-generated.

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.

Keywords:
PDMS devicesoptical trapoptical tweezerssingle cells

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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.