Preparation and sterilization of glassware
Preparation and sterilization of glassware
This lab was derived from: Animal Cell Culture - A Guide to Basic Techniques (5th Edition)
Operation method
Program 11.1 Preparation and Sterilization of Glassware
Materials and Instruments
Disinfectant (hypochlorite) Cleaner Move I. Collection and cleaning of glassware Caveat 1. Keep organic materials outside the oven. Do not use paper tape or packing materials unless you can be sure they will not produce explosive substances when heated. These substances will eventually remain in the oven and produce odors when heated, and some will remain in sterilized glassware. 2. Autoclave at 121°C for 20 min, going through a cycle of pre-vacuum and post-vacuum, either by loosening the screw cap of the bottle or by wrapping the mouth with aluminum foil and then taping it with autoclave tape. After cooling, the loosened cap is then tightened. During autoclaving, the cap should be screwed on very loosely (one turn is sufficient) to allow steam to enter the bottle and to stop the gasket (if any) from being sucked up by the cap and blocking the mouth of the bottle. If the mouth of the bottle is blocked during the autoclave sterilization process, it will result in incomplete sterilization. Unfortunately, during autoclaving there is often a mist which leaves a residue in the bottle as it evaporates. Also the bottle may be contaminated with unsterilized air when it is cooled before being sealed. Dry heat sterilization is relatively good because it allows the bottles to cool directly in the oven before being moved. For more product details, please visit Aladdin Scientific website.
Soaking tubs Test tube brushes Stainless steel baskets Aluminum foil Sterility indicators Sterility indicator tape or plugs Sterility ovens
1. Immediately after use, glassware should be collected in a container that contains a disinfectant (hypochlorite, diluted with a detergent that should contain a minimum of 300 μg/g of chlorine, such as sodium hypochlorite, which is routinely used).
The disinfectant must be collected immediately after use in a detergent (hypochlorite, diluted with a cleaning agent containing a minimum of 300 μg/g chlorine, e.g. sodium hypochlorite for routine use) containing a disinfectant (e.g. 7x or Decon). Glassware should never be allowed to dry out before soaking or washing, otherwise it will be very difficult to clean.
2. Soak in detergent overnight.
3. Rinse:
(a) Early the next morning, rinse glassware by hand or machine with a test tube brush, rinsing 4 times with tap water and then 3 times with deionized water. It is essential to keep the water jet running, otherwise the bottles will have to be filled and emptied each time. Securing many bottles in a basket will speed up the rinsing process.
(b) No detergent is needed for machine cleaning. If a tap is connected, the process can be reduced to two tap water rinses and one deionized water rinse.
4 . After rinsing thoroughly, dry the bottles upside down in a stainless steel basket (holding rinsed glassware and then air drying).
5 . Cool and seal with aluminum foil for storage.
II. Sterilization of glassware
1. Apply small squares of sterility indicator tape to the glassware (this is different from the sterility indicator tape used in autoclaves because of the higher temperatures in the oven. The sterile indicator tape in most autoclaves bakes and the explosive components of the adhesive are left in the oven and in the glassware). Or other indicator labels with the date written on them.
2. place the glassware in an oven (which can reach 160°C with a fan. a recordable thermometer and a bendable probe are desirable) that has a fan that circulates air and is set at 160°C.
3. Ensure that the center of the oven reaches 160°C:
(a) Place a sterility indicator (Alpha Medical for dry heat or steam sterilization, Thermalog indicator for steam sterilization only) in a bottle or other vessel in the center of the oven.
(b) If using a recordable dough meter, place the sensing portion in a bottle or other vessel in the center of the oven.
(c) Do not stack items too tightly, allow space for hot air convection.
4. Cover the oven, check that the temperature knob is at 160°C, seal the oven with tape and hold it for a predetermined time of 1h (or use an automatic lock and automatic recorder).
5. After 1h turn off the power and leave the oven to cool. An automatic timer would be very convenient as it will automatically cut off the power at night and the experiment can be resumed early the next day. This allows cooling to take place in a sterile environment and also minimizes the heat generated during the day.
6 . Sterilized glassware muscle should be used within 24~48h. 