Table of Contents
Overview
The purpose of this page is to provide basic points of the aseptic technique required when doing cell culture and handling all cell culture reagents and materials. Contamination is common with beginners. And every month I hear of yeast infections, bacterial contamination, or something of the sort from the cell culture room.
Must-do techniques
- Active cell lines, media, and all reagents should never be opened outside the BSL hood unless unavoidable.
- EVERYTHING that enters the hood (including your gloved hands, arms) must be sprayed down with 70% EtOH before entering the hood. If there is a possibility that the thing is contaminated, assume it is! If it is irreplaceable and needs to be UV-ed, we have a UV wand!
- The BSL hood you are working with has airflow down and cycles through a HEPA filter, which must be tested every year.
- All cell culture plastics must be in plastic bags and never opened outside of the hood. Tape the encapsulating plastic bags closed after use, and do not remove them from the hood until done so.
- Your exposed arms should never hover over any cell culture reagents, plastics, materials.
- If it can be avoided, nothing should hover over anything.
Incubator notes
- MEFs require 5% CO2, so the incubator is hooked up to a CO2 tank provided by CRLS for ~$2 charge per month. The CO2 tank releases gas based on a program in the hood itself. All you, the user has to do, is connect the tank and set the releasing PSI pressure to 20 PSI.
- At the bottom of the incubator, there should be a tray. This tray should have water in it. Given how dry New Mexico is, this water will provide essential humidity for your cells so the media doesn't evaporate. This phenomenon is especially prevalent in the edge-wells of 96-well plates.
- Use a sharpie to mark what is what on the incubator glass door to stay organized! Don't use the ethanol-proof one!