Single Cell Isolation Using Optical Tweezers

被引:73
|
作者
Keloth, Anusha [1 ]
Anderson, Owen [1 ]
Risbridger, Donald [1 ]
Paterson, Lynn [1 ]
机构
[1] Heriot Watt Univ, Sch Engn & Phys Sci, Inst Biol Chem Biophys & Bioengn, Edinburgh EH14 4AS, Midlothian, Scotland
基金
英国工程与自然科学研究理事会;
关键词
optical tweezers; optical trap; PDMS devices; single cells; ESCHERICHIA-COLI; SYSTEM; WAVELENGTH; GROWTH; MICROSPHERES; TECHNOLOGIES; SEPARATION; FORCES;
D O I
10.3390/mi9090434
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Optical tweezers offer a non-contact method for selecting single cells and translocating them from one microenvironment to another. We have characterized the optical tweezing of yeast S. cerevisiae and can manipulate single cells at 0.41 +/- 0.06 mm/s using a 26.8 +/- 0.1 mW from a 785 nm diode laser. We have fabricated and tested three cell isolation devices; a micropipette, a PDMS chip and a laser machined fused silica chip and we have isolated yeast, single bacteria and cyanobacteria cells. The most effective isolation was achieved in PDMS chips, where single yeast cells were grown and observed for 18 h without contamination. The duration of budding in S. cerevisiae was not affected by the laser parameters used, but the time from tweezing until the first budding event began increased with increasing laser energy (laser power x time). Yeast cells tweezed using 25.0 +/- 0.1 mW for 1 min were viable after isolation. We have constructed a micro-consortium of yeast cells, and a co-culture of yeast and bacteria, using optical tweezers in combination with the PDMS network of channels and isolation chambers, which may impact on both industrial biotechnology and understanding pathogen dynamics.
引用
收藏
页数:21
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