Nano-opto-mechanical characterization of neuron membrane mechanics under cellular growth and differentiation.
We designed and fabricated silicon probe with nanophotonic force sensor to directly stimulate neurons (PC12) and measured its effect on neurite initiation and elongation. A single-layer pitch-variable diffractive nanogratings was fabricated on silicon nitride probe using e-beam lithography, reactive ion etching and wet-etching techniques. The nanogratings consist of flexure folding beams suspended between two parallel cantilevers of known stiffness. The probe displacement, therefore the force, can be measured through grating transmission spectrum. We measured the mechanical membrane characteristics of PC12 cells using the force sensors with displacement range of 10 mum and force sensitivity 8 muN/mum. Young's moduli of 425 +/- 30 Pa are measured with membrane deflection of 1% for PC12 cells cultured on polydimethylsiloxane (PDMS) substrate coated with collagen or laminin in Ham's F-12K medium. In a series of measurements, we have also observed stimulation of directed neurite contraction up to 6 mum on extended probing for a time period of 30 min. This method is applicable to measure central neurons mechanics under subtle tensions for studies on development and morphogenesis. The close synergy between the nano-photonic measurements and neurological verification can improve our understanding of the effect of external conditions on the mechanical properties of cells during growth and differentiation.
Gopal
Ashwini
A
Department of Biomedical Engineering, The University of Texas at Austin, 1 University Station, ENS 12, Austin, TX, 78712-0238, USA.
Luo
Zhiquan
Z
Lee
Jae Young
JY
Kumar
Karthik
K
Li
Bin
B
Hoshino
Kazunori
K
Schmidt
Christine
C
Ho
Paul S
PS
Zhang
Xiaojing
X
D000818:Animals; D001696:Biomechanical Phenomena / Q000379:methods; D018929:Cell Culture Techniques; D002461:Cell Line, Transformed; D002462:Cell Membrane / Q000378:metabolism; D049109:Cell Proliferation; D002478:Cells, Cultured; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D003094:Collagen / Q000737:chemistry; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D004867:Equipment Design; D007797:Laminin / Q000737:chemistry; D040542:Mechanotransduction, Cellular; D036103:Nanotechnology / Q000295:instrumentation / Q000379:methods; D009474:Neurons / Q000502:physiology*; D055095:Optics and Photonics; D016716:PC12 Cells; D051381:Rats; D013314:Stress, Mechanical; D013379:Substrate Specificity; D014159:Transducers
D020099:Coated Materials, Biocompatible; D004129:Dimethylpolysiloxanes; D007797:Laminin; C013830:baysilon; D003094:Collagen