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Nanomanipulation of Single RNA Molecules by Optical Tweezers
Published on: August 20, 2014
Raman tweezers sorting of single microbial cells
Wei E Huang1, Andrew D Ward, Andrew S Whiteley
1Molecular Microbial Ecology, CEH-Oxford, Mansfield Road, Oxford OX1 3SR, UK. Science and Technology Facilities Council, Rutherford Appleton Laboratory, Harwell Science and Innovation Campus, Didcot OX11 0QX, UK.
Environmental Microbiology Reports
|June 15, 2013
Summary
This study introduces Raman tweezers, a novel method combining Raman microspectroscopy and optical trapping to isolate single microbial cells. This technique enables the study of unculturable microorganisms by identifying (13)C uptake via spectral shifts.
Area of Science:
- Microbiology
- Spectroscopy
- Biotechnology
Background:
- Studying unculturable microorganisms is crucial for understanding microbial diversity and function.
- Current methods for isolating single cells often face limitations in efficiency and applicability to unculturable species.
Purpose of the Study:
- To develop and validate a novel technique for the selective isolation and manipulation of single microbial cells based on metabolic activity.
- To demonstrate the potential of Raman tweezers for studying microorganisms that are difficult or impossible to culture using conventional methods.
Main Methods:
- Utilized Raman microspectroscopy to identify microbial cells based on spectral peak shifts, specifically the phenylalanine peak shift indicating (13)C uptake.
- Employed optical trapping with an infrared laser for non-invasive cell capture and manipulation.
- Integrated Raman spectral acquisition with cell sorting and physical separation within capillary tubes.
Main Results:
- Successfully discriminated and isolated single yeast (Saccharomyces cerevisiae) and bacterial cells (Escherichia coli, Pseudomonas fluorescens) exhibiting (13)C uptake.
- Demonstrated cell viability and successful genome amplification from sorted cells, with recovery rates of 61% for yeast and 39% for bacteria.
- Validated the Raman tweezers approach for isolating metabolically active single cells.
Conclusions:
- Raman tweezers offer a powerful new approach for the isolation and study of single, potentially unculturable microorganisms.
- This technique has significant implications for advancing our understanding of microbial ecology and exploring novel microbial functions.
- The method provides a pathway to investigate the vast majority of microbes in natural environments that remain uncharacterized.
