Optical neuronal guidance in three-dimensional matrices.
We demonstrate effective guidance of neurites extending from PC12 cells in a three-dimensional collagen matrix using a focused infrared laser. Processes can be redirected in an arbitrarily chosen direction in the imaging plane in approximately 30 min with an 80% success rate. In addition, the application of the laser beam significantly increases the rate of neurite outgrowth. These results extend previous observations on 2D coated glass coverslips. We find that the morphology of growth cones is very different in 3D than in 2D, and that this difference suggests that the filopodia play a key role in optical guidance. This powerful, flexible, non-contact guidance technique has potentially broad applications in tissues and engineered environments.
Dept. of Physics, Georgetown University, Washington, DC 20057, USA.
Catherine E
CE
Graves
Ryan G
RG
McAllister
William J
WJ
Rosoff
Jeffrey S
JS
Urbach
D000818:Animals; D002454:Cell Differentiation / Q000502:physiology / Q000528:radiation effects; D048430:Cell Shape / Q000502:physiology / Q000528:radiation effects; D003094:Collagen / Q000502:physiology; D020439:Growth Cones / Q000502:physiology / Q000528:radiation effects* / Q000648:ultrastructure; D048091:Guided Tissue Regeneration / Q000295:instrumentation / Q000379:methods*; D007834:Lasers; D009416:Nerve Regeneration / Q000502:physiology / Q000528:radiation effects; D016501:Neurites / Q000502:physiology / Q000528:radiation effects / Q000648:ultrastructure; D055495:Neurogenesis / Q000502:physiology / Q000528:radiation effects; D055095:Optics and Photonics / Q000295:instrumentation / Q000379:methods*; D009924:Organ Culture Techniques; D016716:PC12 Cells; D010775:Photic Stimulation / Q000295:instrumentation / Q000379:methods*; D011554:Pseudopodia / Q000502:physiology / Q000528:radiation effects / Q000648:ultrastructure; D051381:Rats; D023822:Tissue Engineering / Q000379:methods
D003094:Collagen