Therapeutic treatment of spinal cord injuries, brain trauma, stroke, and neurodegenerative diseases will greatly benefit
from the discovery of compounds that enhance neuronal regeneration following injury. We previously demonstrated the
use of femtosecond laser microsurgery to induce precise and reproducible neural injury in C. elegans, and have
developed microfluidic on-chip technologies that allow automated and rapid manipulation, orientation, and non-invasive
immobilization of animals for sub-cellular resolution two-photon imaging and femtosecond-laser nanosurgery. These
technologies include microfluidic whole-animal sorters, as well as integrated chips containing multiple addressable
incubation chambers for exposure of individual animals to compounds and sub-cellular time-lapse imaging of hundreds
of animals on a single chip. Our technologies can be used for a variety of highly sophisticated in vivo high-throughput
compound and genetic screens, and we performed the first in vivo screen in C. elegans for compounds enhancing
neuronal regrowth following femtosecond microsurgery. The compounds identified interact with a wide variety of
cellular targets, such as cytoskeletal components, vesicle trafficking, and protein kinases that enhance neuronal
regeneration.
In recent years, the advantages of using small invertebrate animals as model systems for human disease have become
increasingly apparent and have resulted in three Nobel Prizes in medicine or chemistry during the last six years for
studies conducted on the nematode Caenorhabditis elegans (C. elegans). The availability of a wide array of species-specific
genetic techniques, along with the transparency of the worm and its ability to grow in minute volumes make C.
elegans an extremely powerful model organism. We present a suite of technologies for complex high-throughput whole-animal
genetic and drug screens. We demonstrate a high-speed microfluidic sorter that can isolate and immobilize C.
elegans in a well-defined geometry, an integrated chip containing individually addressable screening chambers for
incubation and exposure of individual animals to biochemical compounds, and a device for delivery of compound
libraries in standard multiwell plates to microfluidic devices. The immobilization stability obtained by these devices is
comparable to that of chemical anesthesia and the immobilization process does not affect lifespan, progeny production,
or other aspects of animal health. The high-stability enables the use of a variety of key optical techniques. We use this to
demonstrate femtosecond-laser nanosurgery and three-dimensional multiphoton microscopy. Used alone or in various
combinations these devices facilitate a variety of high-throughput assays using whole animals, including mutagenesis
and RNAi and drug screens at subcellular resolution, as well as high-throughput high-precision manipulations such as
femtosecond-laser nanosurgery for large-scale in vivo neural degeneration and regeneration studies.
Access to the requested content is limited to institutions that have purchased or subscribe to SPIE eBooks.
You are receiving this notice because your organization may not have SPIE eBooks access.*
*Shibboleth/Open Athens users─please
sign in
to access your institution's subscriptions.
To obtain this item, you may purchase the complete book in print or electronic format on
SPIE.org.
INSTITUTIONAL Select your institution to access the SPIE Digital Library.
PERSONAL Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.