The Journal of Neuroscience, June 7, 2006, ():
Microtargeted Gene Silencing and Ectopic Expression in Live Embryos Using Biolistic Delivery with a Pneumatic Capillary Gun
J. Neurosci. Shefi et al.
26: 6119
Supplemental data
Files in this Data Supplement:
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Supplemental material
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FIGURE LEGENDS
Supplemental Figure 1: Schematic drawing showing the pneumatic capillary gun with a head attached to it mounted on a micromanipulator. Arrows indicate the flow of He.
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Supplemental Figure 2: Transfection and staining of mammalian cells cultured in a Petri dish using particles coated with a plasmid encoding GFP and with a lipophilic dye. A and B, Fluorescence image and a superposition of brightfield and fluorescence images of a cell culture ~24 hr after receiving a shot of particles coated with plasmid DNA encoding GFP. The black dots in panel B are gold particles. The cell layer is nearly confluent and is not visibly damaged by the shot. Expression of GFP is clearly seen in 5 cells. The images are taken with a 10× objective, scale bar 100 μm. C and D, Magnified images of a portion of the area shown in panels A and B, with two transfected cells. Fluorescence image in C and a superposition of brightfield and fluorescence images in D. In addition to the cell bodies, expression of GFP is clearly seen in processes of the cells. Images are taken with a 50× objective, scale bar 10 μm. E and F, Fluorescence image and a superposition of brightfield and fluorescence images of a cell culture after a shot of particles coated with a red fluorescent lipophilic dye (DiI). The number of gold particles (black dots) is much smaller than in panels A-B, and just as in panels A-B, the cell layer is nearly confluent and undamaged. The images are taken with a 10× objective, scale bar 100 μm. G and H, Magnified images of a portion of the area shown in panels E and F , fluorescence in G and a superposition of brightfield and fluorescence in H, with three fluorescently stained cells. All fluorescent cells have gold particles inside. The images are taken with a 50× objective, scale bar 10 μm.
