After two months using Sf9 cells I got a bacterial contamination. I checked my medium and it is fine.
I worked with Sf21 and S2 cells and rarely had contamination. What type of cell medium do you use? Do you supply it with serum and antibiotics? I used penicillin.
The only option you have is to discard those plates and start again with a new vial of cells. But first check if the contamination is in your media: just put a plate with fresh medium and no cells in the incubator. Sometimes, even very experienced people get bacterial contamination in their cultures.
How are you growing your cells (shaker flask or T-flask), each has it's own requirements for keeping the culture inside sterile. But either way you have to move them in and out of the hood, and when they are out of the hood they can get contaminated on the surface, then if you do not spray them down with 70% isopropanol before returning them to the hood the bacteria can bounce around in the hood while your flask is open. Also, does anyone else use the hood? Do you spray down the hood working surface with 70% isopropanol before using? Does the hood have a UV lamp that is still effective?
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