4. Secure dispense tips into desired placement slot on dispense tip head using provided Allen wrench.     
    a. For best results, position tips very close to the membrane surface, without direct contact (unless contact is desired).

    i. Actual height is dependent on membrane type and thickness.
    b. Tips positioned too high may result in liquid droplet formation or uneven lines. 
    c. To test positioning, place membrane on dispense table and initiate switch to cause table to move. Drag on membrane will be apparent.
    i. Once desired height is established, it may be helpful to mark position on metal post of dispense tip with a permanent marker.
5. Once dispense reagents have been prepared, remove air bubbles prior to use in ALFRD (i.e. quick centrifugation, nitrogen air purge).

    a. A volume of 200 µl or greater is recommended.

    b. When solution is running out during dispensing, lines will appear thinner and may produce varying results. 6. Draw up each solution into a syringe, taking care to minimize air bubbles.

7. Attach tubing to blunt syringe needle and secure filled syringes on syringe pump according to manufacturer’s instructions.

8. Turn on syringe pump to prime solution(s) through tubing and dispense. Turn off pump and wipe away residual liquid.

9. Secure membrane onto ALFRD dispense table using magnets.

10. Turn on syringe pump, followed immediately by dispense table switch.

11. Once dispense tips have reached the end of the table and stopped, turn off the pump.

12. Remove membrane from table.

13. Return dispense tips to their original position by reversing the table switch.

14. Wipe off any residual liquid from table.

15. Repeat for each membrane as necessary.