High-throughput optofluidic system for the laser microsurgery of oocytes.

This study combines microfluidics with optical microablation in a microscopy system that allows for high-throughput manipulation of oocytes, automated media exchange, and long-term oocyte observation. The microfluidic component of the system transports oocytes from an inlet port into multiple flow channels. Within each channel, oocytes are confined against a microfluidic barrier using a steady fluid flow provided by an external computer-controlled syringe pump. This allows for easy media replacement without disturbing the oocyte location. The microfluidic and optical-laser microbeam ablation capabilities of the system were validated using surf clam (Spisula solidissima) oocytes that were immobilized in order to permit ablation of the 5 μm diameter nucleolinus within the oocyte nucleolus. Oocytes were the followed and assayed for polar body ejection.

[1]  M. Meseguer,et al.  Oxygen consumption is a quality marker for human oocyte competence conditioned by ovarian stimulation regimens. , 2011, Fertility and sterility.

[2]  M. Pinot,et al.  Towards high throughput production of artificial egg oocytes using microfluidics. , 2011, Lab on a chip.

[3]  J. J. Henry,et al.  Rediscovery of the nucleolinus, a dynamic RNA-rich organelle associated with the nucleolus, spindle, and centrosomes , 2010, Proceedings of the National Academy of Sciences.

[4]  Yu Sun,et al.  In-situ mechanical characterization of mouse oocytes using a cell holding device , 2010, 2010 IEEE 23rd International Conference on Micro Electro Mechanical Systems (MEMS).

[5]  Jared P. Stephens,et al.  Comparative analysis of different laser systems to study cellular responses to DNA damage in mammalian cells , 2009, Nucleic acids research.

[6]  Vasan Venugopalan,et al.  Examination of laser microbeam cell lysis in a PDMS microfluidic channel using time-resolved imaging. , 2008, Lab on a chip.

[7]  G. Whitesides.  The origins and the future of microfluidics , 2006, Nature.

[8]  D. Psaltis,et al.  Developing optofluidic technology through the fusion of microfluidics and optics , 2006, Nature.

[9]  Michael W Berns,et al.  Internet‐based robotic laser scissors and tweezers microscopy , 2005, Microscopy research and technique.

[10]  G. Whitesides,et al.  Rapid Prototyping of Microfluidic Systems in Poly(dimethylsiloxane). , 1998, Analytical chemistry.

[11]  Bruce J. Tromberg,et al.  Optical determination of motility forces in human spermatozoa with laser tweezers , 1996, European Conference on Biomedical Optics.

[12]  M W Berns,et al.  Determination of motility forces of human spermatozoa using an 800 nm optical trap. , 1996, Cellular and molecular biology.

[13]  R Yanagimachi,et al.  Intracytoplasmic sperm injection in the mouse. , 1995, Biology of reproduction.

[14]  P. Devroey,et al.  Pregnancies after intracytoplasmic injection of single spermatozoon into an oocyte , 1992, The Lancet.

[15]  G. Dessev,et al.  Effect of Calcium on the Stability of the Vitelline Envelope of Surf Clam Oocytes. , 1990, The Biological bulletin.

[16]  E. Kinney.  Primer of Biostatistics , 1987 .

[17]  B. Horwitz.  RATES OF OXYGEN CONSUMPTION OF FERTILIZED AND UNFERTILIZED ASTERIAS, ARBACIA, AND SPISULA EGGS. , 1965, Experimental cell research.

[18]  P. Wildy,et al.  CYTOCHEMICAL STUDIES OF THE NUCLEOPROTEINS OF HELA CELLS INFECTED WITH HERPES VIRUS , 1963, The Journal of cell biology.

[19]  R. Allen.  FERTILIZATION AND ARTIFICIAL ACTIVATION IN THE EGG OF THE SURF-CLAM, SPISULA SOLIDISSIMA , 1953 .

[20]  W. Kinsey.  Analysis of signaling pathways in zebrafish development by microinjection. , 2009, Methods in molecular biology.

[21]  M. Tengowski.  Microscopic techniques for studying sperm-oocyte interaction during fertilization and early embryonic development. , 2004, Methods in molecular biology.

[22]  G. Whitesides,et al.  Fabrication of microfluidic systems in poly(dimethylsiloxane) , 2000, Electrophoresis.