Microfluidic immobilization of physiologically active Caenorhabditis elegans.
We present a protocol for building and operating a microfluidic device for mechanical immobilization of Caenorhabditis elegans in its physiologically active state. The system can be used for in vivo imaging of dynamic cellular processes such as cell division and migration, degeneration, aging and regeneration, as well as for laser microsurgery, Ca2+ imaging and three-dimensional microscopy. The device linearly orients C. elegans, and then completely restrains its motion by pressing a flexible membrane against the animal. This technique does not involve any potentially harmful anesthetics, gases or cooling procedures. The system can be installed on any microscope and operated using only one syringe and one external valve, making it accessible to most laboratories. The device fabrication begins by patterning photoresist structures on silicon wafers, which are then used to mold features in elastomeric layers that are thermally bonded to form the device. The system can be assembled within 3 d.
Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
Cody L
CL
Gilleland
Christopher B
CB
Rohde
Fei
F
Zeng
Mehmet Fatih
MF
Yanik
D000818:Animals; D017173:Caenorhabditis elegans / Q000502:physiology*; D003952:Diagnostic Imaging / Q000295:instrumentation / Q000379:methods; D044085:Microfluidics / Q000295:instrumentation* / Q000379:methods; D012149:Restraint, Physical / Q000295:instrumentation* / Q000379:methods