$page[title] = "Growth and Alignment of Protein Fibers with Glass Rods"; $page[show_updated] = 1; ?>
By Michael Sawaya ala Magdalena Ivanova.
You will need:
The glass pipets are 12 cm long. Break few of the pipets into segments approximately 1.5 to 2.5 cm long. Using the tweezers, hold the end of the glass capillary in a flame to seal the end. Turn the capillary around and seal the other end of the tube. Scratch one of the ends of sealed capillary with sandpaper. Stick the non-scratched end in a small chunk of clay. Use the clay to fix the capillary in the petri dish. Place a second capillary 1 to 2 mm away (tip to tip distance) as shown in the attached photo.
Pellet and wash the sample as described below:
The procedure involves first removing any salt that might be present in the prion sample. (Salt diffracts strongly and so must be removed). We do this by pelleting the material in a table top centrifuge (20,000 X g for 5 minutes), resuspending the pellet in water, then pelleting again. The pellet is resuspended in 5 microliters of water then pipeted between two glass rods. The glass rods are fire polished at the tips and then scratched with sand paper to allow the fiber material to stick to the glass. The fibers are ready after a few hours. You should see a fiber stretched between the ends of the glass rods.