Sterilization and filtration with small tandem filters
Sterilization and filtration with small tandem filters
This lab was derived from: Animal Cell Culture - A Guide to Basic Techniques (5th Edition)
Operation method
Option 11.13 Sterilization and Filtration with Small Tandem Filters
Materials and Instruments
Media to be sterilized Move 1. Connect the tubing (silicone tubing, fitted to the peristaltic pump, to the filter connection) to the peristaltic pump (preferably with a bottom switch). For more product details, please visit Aladdin Scientific website.
Tandem filters Culture medium bottles Lids Containers Peristaltic pumps Silicone tubes Racks
2. Insert the upstream end into the medium to be filtered.
3. Unpack the filter and connect it to the peristaltic pump outlet.
4. Clamp the filter (tandem filter with hood) at a suitable height in the rack (fixed filter) so that the bottle in which the medium is to be collected (graduated medium bottle, foil-wrapped, sterilized by dry heat) is just below the filter, the neck of the bottle can be covered and the bottle can be easily removed when full. If desired, a sterile culture bottle can be used first for commissioning, but do not use this bottle to collect the sterile liquid at the end.
5. Turn on the pump and collect about 20 ml in the bottle for commissioning.
6. Remove the commissioning bottle (autoclaved), close the cap and mark it with "1".
7. Remove the aluminum foil from the first medium bottle and place it under the filter bell.
8. Turn on the pump.
9. Fill bottle to 100 ml.
10. Turn pump off.
11. Remove, cap, and number the bottle and replace with a new sterile media bottle.
12. Repeat steps 7 to 11, filling 5 bottles with 100 ml each and collecting the remainder in the last bottle.
13. Fill the upper space of each bottle with 5% CO2 gas.
14. Re-wrap the aluminum foil around the lid and up to the neck of the bottle to ensure dust free storage.
15. Label, date, and sign bottles.
16. Store the bottles containing 100 ml of medium at 4°C.
17. Incubate for one week at 37°C with the first and last bottles capped.
18. Store medium at 4°C and do not use until sterility testing is complete. If contamination is found in the test sample, the entire batch of medium should be re-filtered or discarded.