How to Operate Microfluidizer (100 mL Minimum Sample)

!!!!DO NOT FLOW CLUMPY SAMPLE THROUGH THE SYSTEM!!!!

1. Check if cooling unit (metal cylinder with coil) contains any water.
   If so, loosen clamp and drain.
   Tighten clamp and fill cooling unit with an ice water mixture.

   Depending on how long it takes to process your sample, you may need to drain water and add more ice periodically.

2. If sample port (glass cylinder) is empty, fill to top with water.
   If 70% ethanol is still present, leave it.

3. Turn on unit (white switch on left side), then motor (green button on front).

   Should get very noisy.

4. PRIME SYSTEM

   • Open silver valve.
   • Pull up intensifier (black button on front) to start flow.
   • After 3–4 pumps, push black button to stop.
   • Close valve.

5. CHECK PRESSURE

   • Open black valve.
   • Pull up black button.
   • Adjust to desired pressure with knob on front.

     It should be more than 4000 PSI.

   • Push black button to stop.
   • Close valve.

6. Start flow of water

   • Pull up black button to start.
   • Shortly before liquid reaches the bottom, push black button to stop flow so air doesn't enter system.

7. Add ~100 mL buffer to sample port. Let it flow through.

8. Add your sample to the sample port.

   • Start flow.
   • Let ~5 pumps flow through before collecting your sample (void volume).

9. Pass sample through system as many times as desired.

10. After last sample pass:

    • Add ~100 mL buffer.
    • Collect another 7 pumps to get remaining sample from void volume.

11. CLEAN SYSTEM (Step 1):

    • Fill sample port to top with water.
    • Let flow through main valve.
    • After water has flowed through, repeat with water through the prime valve.

12. CLEAN SYSTEM (Step 2):

    • Add 70% ethanol.
    • Wash through prime valve, then main valve.

13. Add 10 mL 70% ethanol.

    • After 6 pumps through the main valve, stop flow.
    • Turn off system (red button on front, then white switch on side).

14. Loosen cooling unit clamp to drain water.