This lab was derived from: Animal Cell Culture - A Guide to Basic Techniques (5th Edition)

Operation method

Option 11.14 Sterilization and Filtration with Large Tandem Filters

Materials and Instruments

Tandem Filters Media Bottles Lids Containers Peristaltic Pumps Silicone Tubing Racks

Move

1. Place the filter racks (clips and clip holders that protect the filters (unless the filter racks have legs)).

2. Attach the outlet to the receiver bottle (filter receiver container with outlet on the bottom).

3. place the outlet of the filter (e.g., 90 mm membrane and reusable filter holder, or disposable disc or cartridge filter) at a suitable height so that the bottle in which the medium is to be collected (medium bottle, foil-wrapped, sterilized by dry heat method) is located just below the filter, the mouth of the bottle can be covered by a bell jar, 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 solution at the end.

4. Connect the pressure bottle to the inlet of the filter.

5. Slowly pour the culture medium into the pressure bottle (5 ~ 50L).

6. For reusable filters, leave the pump (100 kPa (1 bar, 15 p. s. i.)) on long enough to moisten the filter. Then turn the pump off and tighten the filter holder.

7. Turn the pump on to 100 kPa (15 ^{lb/in2 )} . When the receiver container (filter receiver container with outlet at the bottom) is almost full, open the tubing (silicone pressure-resistant tubing) to the media storage bottle.

8. Remove the bottle from under the filter bellows and cap it.

9. Re-wrap the aluminum foil around the lid (the lid of the culture medium bottle, autoclaved) up to the neck of the bottle to ensure dust-free storage.

10. Label, date, and sign the bottle.

11. Depending on the size and number of bottles collected after filtration, take samples from the bottles at the beginning, middle, and end of filtration and incubate at 37°C for one week to check for contamination.

12. Store at 4°C. Do not use until quality control is complete.

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