title: High-throughput optofluidic system for the laser microsurgery of oocytes.
abstract: 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.
authors.affiliation: University of California, San Diego, Department of Bioengineering, 9500 Gilman Drive, La Jolla, California 92093, USA. cchandsa@ucsd.edu
authors.forename: Charlie
authors.initials: C
authors.lastname: Chandsawangbhuwana
authors.forename: Linda Z
authors.initials: LZ
authors.lastname: Shi
authors.forename: Qingyuan
authors.initials: Q
authors.lastname: Zhu
authors.forename: Mark C
authors.initials: MC
authors.lastname: Alliegro
authors.forename: Michael W
authors.initials: MW
authors.lastname: Berns
mesh_terms: D000818:Animals; D002467:Cell Nucleus; D004867:Equipment Design; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods; D053685:Laser Therapy; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D008866:Microsurgery / Q000295:instrumentation* / Q000379:methods*; D009865:Oocytes / Q000166:cytology*; D055095:Optics and Photonics / Q000295:instrumentation; D015203:Reproducibility of Results; D049890:Spisula / Q000166:cytology