Sterilization of Glassware, Equipments and Culture Media
The thoroughly washed glassware and equipments and carefully prepared culture media are sterilized by heat, steam or millipore filter paper (Table 6.4 ). The sterilized materials are removed when temperature cools down and used according to procedure adopted. For detailed description
see In Vitro Culture Techniques: The Biotechnological Principles.
Table 6.4. Sterilization of glassware and equipments.
Sterilization |
Items |
Autoclaving* |
Apparatus containing glass and silicon tubing, disposable tips for micropipettes, dispensor tubing for pupet, glass bottles with screw caps, magnetic stirrer bars, screw caps, silicon tubing, topper-rubber, silicon, millipore filters, etc. |
Dry heat** |
Glassware, glass coverslips, glass slides, instruments, Pasteur pipettes, pipettes, test tubes, etc. |
*15 lb/in
2 (=121°C) for 20 minutes; **160°C/ 1 hour.
Table 6.5. Sterilization of liquid by different methods.
Sterilization |
Storage |
Solution |
Autoclave* |
Room temperature |
Agar, antibiotics, EDTA, glucose, glycerol, HEPES, lactalbumin hydrolysate, NaHCO3 solution, NaOH solution, phenol red, salt solution (with glucose), tryptose, water |
Autoclave |
4°C |
Methocel |
Filter** |
Room temperature |
Glucose solution, HC1, NaHCO3 solution, NaOH solution, |
Filter |
-20°C |
Collagenase, antibodies, glutamine, sodium pyruvate solution, transferrin, trypsin, vitamines |
Filter-stacked filters |
4°C |
Bovine serum albumin |
Filter-stacked filter |
-20°C |
Serum |
Steam (30 min at 100°C) |
4°C |
Carboxylmethylcellulose |
*15lb/in
2 (=121°C) for 30 minutes, **0.2 mm pore size filter.