Keeping the culture tube at an angle, insert the loop into the broth and remove a loopful of inoculum.
Partially lift the lid of the plate culture and open it just enough to insert the inoculation loop. Do not completely open the lid and expose the surface to the air.
Pick up the tube of sterile nutrient broth with your free hand, carefully remove the cap (cotton plug) with your little finger of the hand holding the loop, and flame the mouth of the tube.
Flame the mouth of the tube. Keeping the culture tube at an angle, insert the loop into the broth and remove a loopful of inoculum. Flame the mouth of the tube again and recap the tube. Place the culture tube back on the test tube rack.
To cool your loop or needle quickly, place it on a section of agar that is uninoculated or is at least different from the area from which you will remove cells.
Almost same but only difference is: you take the sample plate and inoculate into other plates for checking different variety of microbes in the sampled area.
Choose an isolated colony and scrape off a very small amount of culture with the loop. Be careful not to gouge into the agar with the loop as you pick up the organisms Close the lid immediately once you have picked up the organisms and turn the plate upside down on the work surface.
Good aseptic technique is essential for successful long-term cell and tissue cultures. Strict adherence to these principles and practices provides the following benefits for your cultures (Freshney, 2002): Protects the cell line from microbial and cellular cross contamination.
Aseptic technique is a set of principles and practices used by cell culture workers to reduce the potential of unwanted microorganisms or other cell lines from being introduced into: Cell cultures. Sterile solutions and supplies. And, most importantly, the technician and coworkers.
Cell culture contamination is very difficult to avoid entirely, especially in a busy laboratory. However, careful aseptic technique, appropriate caution and training can significantly reduce the risk of contamination within your cell culture facility. All personnel should receive training and demonstrate proficiency in the laboratory practices for conducting and supporting sterile procedures and facility safeguards. Education and technical training programs need to be ongoing endeavors to ensure the quality of work in the long term (Coecke; 2007). Additionally, aseptic technique must be supported by good laboratory design, selection of proper equipment and facility safeguards to protect laboratory workers and to reliably achieve successful sterile manipulation and processing of cell cultures. Bionique is here to help, please feel free to contact the laboratory with any questions.
Avoid sharing bottles of media or other solutions with coworkers. Cross contamination and lack of accountability arise from sharing with others.
This important step reduces both the possibility of cross contamination with another cell line and limits the spread of contamination if the bottle of medium becomes contaminated.
If clean-up of the contaminated culture is attempted, then any work with this culture should be reserved to the very end of the day to minimize transfer of the contamination.
The drop of liquid that usually remains on the lip of the vessel can easily form a liquid bridge between the nonsterile outside and sterile inside of the vessel . This allows microorganisms from the outside to enter and contaminate the vessel and its contents, especially when pouring a second time.