====== Media change ======

===== General media change procedure =====


  - Put the media you will use around 30 min in advance to room temperature
  - 10 min before it will be used – warm it in the beads bath 
  - Spray a falcon with media with EtOH before placing it under the hood, avoid erasing the labels. If erased – name it the same way as it was named. 
  - Place the media to the right side of the hood (“clean-to-dirty” rule). Work with the plates in the middle of the hood, where the airflow is sterile
  - Take out a plate from the bottom incubator, wipe it with EtOH before placing under the hood. Careful to not erase the labels, if erased – add names back
  - Pay attention to which type of media is written on the plates, change media according to labels. It’s very important to not mix it up 
  - Pre-open the media falcons
  - Open the plate and place the cover “face down”
  - Avoid going with your hands above the plate or opened media
  - Aspirate the old media with the pump. Glass pipette should not touch walls of wells and the bottom of the well. Try to hold it 90 degrees to the plate’s surface
  - Minimize the time cells stay without media. They are very sensitive and die easily 
  - Add pre-warmed media with the plastic serological pipette. Don’t re-use the pipette, add media in a steady slow flow to the wall of a well. Try to avoid drops (that will hurt cells). 
  - Close the lid and place the plate back to the incubator. 
  - Proceed with the next one. 

Don’t work with more than 2 plates at the same time. It’s not safe to put plates on top of each other, they will occupy quite some space, which will reduce the area you can sterilize and work on. The cells are also very sensitive, they easily “get shocked” by lower than 37 temperatures. The time outside the incubator needs to be minimized.