Computational protocol: Miniaturized Embryo Array for Automated Trapping, Immobilization and Microperfusion of Zebrafish Embryos

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Protocol publication

[…] The microfluidic chip was designed and modeled using CorelDraw X4 (Corel Corporation, Ontario, Canada) and SolidWorks 2011 (Dassault Systemes SolidWorks Corp, Concord, MA, USA) CAD packages. Subsequent prototyping and fabrication was performed using a non-contact, 30 W CO2 laser cutting system equipped with a High Power Density Focusing Optics (HPDFO)™ (Universal Laser Systems, Scottsdale, AZ, USA). The master shape was laser cut in 1.5 mm thick poly-methyl methacrylate sheet (PMMA/Acrylic; PSP Plastics Ltd, Auckland, New Zealand) to form the integrated channel and trapping region structure (). The master was then thermally bonded to a 25×75×2 mm PMMA sheet at 110°C for up to 2 hours in a fan assisted oven. A uniform mechanical force was applied using two mini C-clamps to provide equal force distribution during PMMA layer bonding. The bonded PMMA master was then used for replica molding in poly(dimethylsiloxane) (PDMS; Sylgard 184; DowCorning Corp, Midland, MI, USA), as described earlier . Briefly, the PDMS was mixed at a 10∶1 (w/w) ratio of elastomer base to curing agent and degassed at 40 Torr to remove any residual air bubbles. PDMS was then poured on a PMMA master to achieve approximately 5 mm thickness and cured thermally at 80°C for up to 1 hour. Cured PDMS devices were mechanically diced and bonded to the glass microscope slides using oxygen plasma surface activation. Tubing interconects were manually bored using an appropriate stainless steel punch hole, as described earlier , . [...] Data analysis and presentation was performed using the LAS (Leica Microsystems); ImageJ (freely available at http://rsb.info.nih.gov/ij/web page); GraphPad Prism (GraphPad Software, Inc. CA, USA); SolidWorks 2011 (Dassault Systemes SolidWorks Corp) and Fluent 6.3 (Fluent) software. The Student’s t-test was applied for comparison between groups using MS Excel (Microsoft, USA) and GraphPad Prism (GraphPad Software) with significance set at p<0.05.All control measurements are provided in detail in the Figure legends, where appropriate, but in general involved making direct comparisons between the chip-based devices and static 24-well microtiter plates or 60 mm Petri dishes (Nalge Nunc Inc, NY, USA). […]

Pipeline specifications

Software tools CorelDraw, ImageJ
Applications Miscellaneous, Microscopic phenotype analysis
Organisms Danio rerio, Homo sapiens
Diseases Drug-Related Side Effects and Adverse Reactions