Spinner Bottle Bioreactor

SOP for Bellco Micro-Carrier Spinner Flasks

  1. Wash all parts with a soft cloth, warm water and Liquinox detergent. Rinse several times with tap water. Rinse several times, inside and out, with deionized water. Air dry.
  2. Gather all parts: glass spinner flask body; sidearm caps; top cap; compression fitting body; compression fitting retainer nut; cap liner (red rubber); shaft lock consisting of cap nut, washer, and o-ring; glass impeller shaft; teflon impeller shaft containing a slot for the impeller and a hole for the magnet; silicone plug; magnet; and impeller with paddle blades.
  3. Assemble Spinner Flask:

    1. Pick up teflon impeller shaft containing magnet holder.
    2. Insert glass impeller shaft (unknobbed end) into the hole at the bottom of the teflon shaft containing the magnet holder and push up until the glass impeller shaft until the knob catches. (It should spin freely.)
    3. Insert silicone plug into the hole at the bottom of the teflon shaft containing the magnetic holder.
    4. Insert impeller with paddle blades into slot at the bottom of the teflon shaft.
    5. Insert magnet through magnet holder on the bottom of the teflon shaft (and through the impeller).
    6. Check to make sure that magnet is equally distributed on both sides of impeller.
    7. Pick up top cap and insert cap liner (red rubber).
    8. Push long threaded portion of compression fitting body up through the hole in the bottom of the top cap until the integral compression fitting nut rests firmly on red rubber cap liner.

    9. Lock compression fitting in place from the top of the cap with retainer nut.
    10. Gently slide the glass impeller shaft through the cap from the bottom and up through the compression fitting on the top cap.
    11. Slide the o-ring onto the glass impeller shaft.
    12. Insert impeller assembly into glass spinner flask body by gently bending the paddle blades on the impeller so they will fit through the center neck opening of the glass spinner flask body.
    13. Screw top cap onto spinner flask.
    14. Adjust the height of impeller assembly by sliding the glass shaft up or down just until the paddle blades and teflon shaft do not touch the floor of the glass spinner flask.
    15. Test the behavior of the impeller assembly by placing on the 9-place magnetic stirrer in the dual CO2 Incubator.
    16. Repeat steps n and o until the impeller spins properly.
    17. Slide the washer onto the glass impeller shaft and on top of the o-ring with the flat surface of the washer touching the o-ring and the beveled surface up.
    18. Slide the cap nut onto the glass impeller shaft on top of the washer and tighten.
    19. Screw on side arm cap(s).
    20. The spinner flask is now ready to use.
    21. Autoclave before or after filling with media (remember to loosen side arm caps before autoclaving).

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