Stimulating Neuronal Cell Differentiation Using Gold Nanorods

Abstract

Source: Paviolo., et al. Gold nanorod-assisted optical stimulation of neuronal cells. J. Vis. Exp. (2015).

This video demonstrates a technique to promote neuronal differentiation by exposing nerve cells to gold nanorods and laser light, which forms cell projections supported by the βIII-tubulin. Staining and observing tubulin and the cell nucleus with epifluorescence microscopy reveals the cell outgrowths, indicating the nanorods' efficacy in neuronal differentiation.

Protocol

1. NG108-15 Neuronal Cell Line Culture and Differentiation

1. For the cell culture medium, prepare 500 ml of sterile Dulbecco's modified Eagle medium (DMEM) containing 10% (w/v) fetal calf serum (FCS), 1% (w/v) L-glutamine, 1% (w/v) penicillin/streptomycin and 0.5% (w/v) amphotericin B.
   Note: Supplements can be aliquoted, stored at -20 °C, and added to the media on the day required. Cell culture medium can be refrigerated in a sterile condition for a maximum of 1 month.
2. For the cell differentiation medium, prepare 50 ml of sterile DMEM containing 1% (w/v) L-glutamine, 1% (w/v) penicillin/streptomycin, and 0.5% (w/v) amph....

Results

Figure 1. Optical fiber experimental setup (A) and average laser irradiances as a function of the laser power for a laser beam of area equals to 0.4 mm2 (B). Beam parameters are (A): the half-angle of the maximum cone of light exiting the fiber (θ), t.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Au NR	Sigma Aldrich	716812
NG108-15	Sigma Aldrich	8811230
DMEM	Sigma Aldrich	D6546
FCS	Life Technologies	10100147
L-glutamine	Sigma Aldrich	G7513
Penicillin/streptomycin	Life Technologies	15140122
Amphotericin B	Life Technologies	15290018
Formaldehyde	Sigma Aldrich	F8775
Triton X-100	BDH	T8532
BSA	Sigma Aldrich	A2058
Anti-βIII-tubulin	Promega	G7121
TRITC-conjugated anti-mouse IgG antibody	Sigma Aldrich	T5393
DAPI	Invitrogen	D1306
UV-Vis spectrometer	Varian Medical Systems Inc.	Cary 50 Bio
Mini centrifuge	Eppendorf	Mini Spin
Sonic bath	Unisonics Australia	FPX 10D
Cell culture incubator	Kendro	Hera Cell 150
Cell culture centrifuge	Hettich	Rotofix 32A
Laser diode	Optotech	780 nm single mode fibre - coupled LD
Optical fiber	Thorlabs	780 HP
Power meter	Coherent	Laser Check
ImageJ	http://rsb.info.nih.gov/ij/index.html
Epifluorescent microscope	Axon Instruments	ImageX-press 5000A
PBS
Polystyrene 96 well plate
CO2
t75 flasks