Abstract
Baker’s yeast (Saccharomyces cerevisiae) represents a very popular single-celled eukaryotic model organism which has been studied extensively by various methods and whose genome has been completely sequenced. It was also among the first living organisms that were manipulated by optical tweezers and it is currently a frequent subject of optical micromanipulation experiments. We built a microfluidic system for optical trapping experiments with individual cells and used it for the assessment of cell tolerance to phototoxic stress. Using optical tweezers with the wavelength of 1064 nm, we trapped individual Saccharomyces cerevisiae cells for 15 min and, subsequently, observed their stress response in specially designed microfluidic chambers over time periods of several hours by time-lapse video-microscopy. We determined the time between successive bud formations after the exposure to the trapping light, took account of damaged cells, and calculated the population doubling period and cell areas for increasing trapping power at a constant trapping time. Our approach represents an attractive, versatile microfluidic platform for quantitative optical trapping experiments with living cells. We demonstrate its application potential by assessing the limits for safe, non-invasive optical trapping of Saccharomyces cerevisiae with infrared laser light.
Highlights
Baker’s yeast (Saccharomyces cerevisiae) and its ability of fermentation gave rise to the first microbial biotechnology ever known to human kind probably around 10,000 years ago [1]
The values of generation time (GT), M and cell area index (CAI) obtained from our experiments are summarized in Table 1, which includes a section presenting the data of Aabo et al [20] for direct quantitative comparison
We have found the cells of S. cerevisiae highly tolerant to optical trapping powers up laser power Pc
Summary
Baker’s yeast (Saccharomyces cerevisiae) and its ability of fermentation gave rise to the first microbial biotechnology ever known to human kind probably around 10,000 years ago [1]. From those prehistoric times, the importance of this unicellular fungus has grown ever larger. Baker’s yeast is the largest source of our knowledge of eukaryotic cellular machinery down to the level of individual protein interactions [3] It was among the first living microorganisms manipulated by a focused laser beam soon after the demonstration of single-beam optical trapping by Arthur Ashkin [4]
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