Keys to Success
Develop and follow cleaning protocols.
Use care in handling glassware. Most breakage occurs during the cleaning process.
Rinse all glassware as soon as possible after use.
Stockpile soiled items in water containing a disinfectant or cleanser to avoid making soiled items harder to clean.
To decontaminate ware, autoclave contaminated items in water containing
cleanser.
Use 7-X® or 7-X O-matic® detergent for
thorough cleaning of all laboratory glassware.
Clean glass by scrubbing with a brush. Periodically inspect brushes for wear to avoid scratching glass.
Disassemble spinner assemblies, compression fittings and all multi-part assemblies for cleaning and to avoid the trapping of disinfectants and cleansers. Failure to do so may result in release of these compounds during use with possible detrimental effects to cultures or assays.
Follow the appropriate cleaning directions for silicone, plastic, rubber, stainless steel and other materials.
Thoroughly rinse items in tap water, followed by distilled or deionized water. Even the smallest amounts of cleansers, disinfectants or acids can affect the final performance of the products. Rigorously follow your rinsing protocol.
Dry glassware on racks or peg boards and inspect after drying. If glassware is hazy, has a film or blotches are evident, then additional cleaning is required before use. Inspect glassware for scratches, chips and cracks.
Do not use scratched, chipped or cracked glassware for centrifugation, pressure, vacuum, heating or freezing. Breakage may result.
Small chips can be flame polished to remove sharp edges. Repair or discard broken glassware. Please contact Bellco to request a quotation to repair glassware. Store ware in cabinets upside down or with openings covered to prevent dust contamination during storage.
Cleaning Pipettes:
Place pipettes in a pipette jar or decontamination can with the tip down, as soon as possible after use. Avoid dropping pipettes into jar as this may damage the tips. Water level in the jar should cover the entire pipette if possible. Place disinfectant or detergent in jar to help reduce soiling during soaking. Decontaminate pipettes in container with water and cleanser being sure to fully immerse pipettes Autoclave on wet cycle.
To remove cotton plugs, mount flexible hose adapter on faucet. Attach three inch segment of tubing to adapter and secure with hose clamp. With water slowly flowing, insert pipette tip into open end of hose. Water pressure will flush cotton plug from the mouthpiece and will rinse pipette. Soak pipettes in 7-X® detergent, acid wash or place in an automatic washer to clean. Rinse thoroughly in tap water followed by rinsing in distilled or deionized water. Thoroughly dry the pipettes.
Use a Bellco Pipette Plugger to stuff cotton into mouthpieces and load into Bellco pipette cans. Dry heat sterilize at 160°C (320°F) for at least two hours. If pipettes are steam sterilized, then dry thoroughly in a drying oven before use.
Ashing Glassware
Ashing reduces all organic compounds to a mineral and carbon ash. To ash, place suitable ware in an oven set at 450°C (842°F) for at least three hours. Allow ware to cool. Rinse and clean as stated above.
Chromic Acid Wash:
Follow all normal safety precautions when using concentrated acids and acid solutions. Acids can severely burn the skin. Dispose of all acids properly. A chromic acid wash has hydrolytic and oxidative capabilities for the chemical decomposition of biological molecules. The acid may also dissolve mineral deposits. To prepare a chromic acid wash, mix 20g of sodium or potassium chromate with sufficient distilled H20 to make a paste of chromate salt. Add 300ml of concentrated sulfuric acid. Increase the proportions to make larger amounts. Use the mixture until it turns green in color. It may be necessary to let heavily soiled items soak overnight.
Mineral Deposits:
Soak glassware in concentrated sulfuric, hydrochloric or nitric acid. Follow all precautions for the handling of these solutions. Hydrochloric acid diluted to 1M in water is often used to remove mineral scales built up in water distillation apparatus. Allow mineral to dissolve and rinse throughly in water. Nitric acid is useful for cleaning stainless steel and removing rust stains.
Cleaning Rubber and Silicone Component:
Silicone and rubber components may absorb cleaners and disinfectants that may leak out during use. It is best not to use these cleansers or solutions to clean these products. Place silicone and rubber components in distilled water after use. Let soak. Rinse exhaustively with tap water. Be sure tubing is completely flushed. Place items in a container of distilled water and put through a wet autoclave cycle or boil. Rinse items in distilled or deionized water and inspect. Discard stained or soiled items.
Preparation of Glassware for Tissue Culture:
The following treatments have been effective in improving cell attachment and growth under certain conditions. For example, attachment of Vero cells is enhanced by sodium hydroxide treatment; duck embryo Fibroblasts attach more readily to sodium carbonate treated glassware. However, build up may occur, so it is best not to use these procedures without an occasional acid wash. Fill roller or tissue culture bottles with the indicated solutions. Coverslips and microscope slides can be treated in beakers containing the solutions. Glass is a weak ion exchanger and thus the surface charge can be modified by exposure to acids and bases. Since most cells are negatively charged, modification of the glass surface to a positive charge usually results in increased cell attachment.
Sodium Hydroxide Treatment:
Fill clean container with a solution of NaOH (0.4g/Liter)(1000ml).
Cover container neck with aluminum foil and autoclave for 45 minutes, liquid cycle.
Empty container, fill once with distilled deionized H20, rinse and drain.
Oven sterilize dry items at least four hours at 177°C (350°F).
Sodium Carbonate Treatment:
Fill clean container with a 0.1% solution of Na2CO3 (1g/Liter)
Cover container with aluminum foil. Autoclave standing up in a shallow pan of water for 45 minutes, liquid setting.
Let the container cool. Empty and rinse once with distilled water. Then drain.
Oven sterilize for at least four hours at 177°C (350°F).
Collagen Treatment:
Prepare collagen. Dissolve 50mg collagen in 83ml glacial acetic acid that has been diluted 1/1000 in sterile, distilled, deionized water. Stir overnight in cold room to dissolve. Spin at 1500 RPM for 15 minutes. Remove clear supernatant and discard pellet. Freeze filter sterilized supernatant at -20°C (-24°F) in aliquots of 10ml. Acid wash the container overnight. Rinse by filling container completely four times with deionized water and once with sterile distilled deionized water. Add 10ml sterile collagen to container. Roll bottle or let stand overnight. Decant collagen. Add 50 ml sterile Tris Buffer to container. Roll bottle or let stand until needed.
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