Aseptic Techniques- Cell Culture and Maintenance
Culturing and maintaining the cells are the basic techniques of any cell biological experiment. With a few of the following tips, one can surely ensure a contamination-free culture dish.
1. Make sure your plates, pipettes, pipette gun, gloves are sterile. Use ethanol. 70% ethanol is decontaminant.
It is best if you have 2 different labcoats (only if you could afford) for open bench experiments and cell culture. If your lab doesn't have two coats to offer, that is fine. You could use the same coat for both. BUT make sure NEVER to touch your coat with the gloves on that you will be using for cell culture.
A cheaper option is change gloves when doing cell culture. It is best to change gloves if your doing an DNA or RNA extraction OR switching to cell culture experiments. Gloves aren't that expensive anyways.
Do not note that you can reuse the gloves for their respective purposes once you are done with one experiment. Eg. If you have been doing some Westerns. During the blocking, you decided to trypsinise your cells. Then, change gloves before cell culture. While waiting for cells to incubate in trypsin, you may change back to the gloves used for Westerns to continue with it. When it is time to attend to your cells, change back to the gloves for cell culture.
Alright I know it may sound confusing 😕, until you try it. The key idea is to have two separate gloves when you are multi-tasking between open bench and cell culture experiments.
2. Before you step in to put your items into the BSC. Rub your gloves with ethanol. Make sure after that, you do not touch your lab coat or anything that will not go into the BSC.
Spray ethanol on the BSC table/bench. Wipe it clean. If you have an aspirator, check if it is working and clean with ethanol before you put any items inside. You may choose to UV the BSC as well.
Pick up the items one by one. Spray ethanol throughout. Wipe once and spray again before putting the item into BSC.
3. UV: one may choose to UV before or after (or both times) performing cell culture experiments. UV is another layer of protection against any microbial source of contamination.
NEVER UV with: your healthy cells, media containing FBS or any drugs/ reagents.
FBS, drugs or some reagents contain proteins. UV may rupture their structure, causing them to lose their activity.
For more tips and visual explanation click the link to get more useful information.
The Cell Culture Basics handbook from Gibco and videos can help your lab attain reproducible results every day. Cell culture is critical to