Bell:Flow-through cleaning of the rotor: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
mNo edit summary |
mNo edit summary |
||
| Line 4: | Line 4: | ||
# Flush through the system 10L of distilled water to remove all traces of buffer and cells. | # Flush through the system 10L of distilled water to remove all traces of buffer and cells. | ||
# Make sure no cells are left at the distal end of the chamber (cells often pellet at that location). If there are still cells, flush through more water with a higher pump rate. If that doesn't work, follow the protocol [[Bell:Cleaning the elutriation chamber | Cleaning the elutriation chamber]]. | # Make sure no cells are left at the distal end of the chamber (cells often pellet at that location). If there are still cells, flush through more water with a higher pump rate. If that doesn't work, follow the protocol [[Bell:Cleaning the elutriation chamber | Cleaning the elutriation chamber]]. | ||
# Now make sure that: | |||
# | |||
## the system is completely filled with water with no air bubbles; | ## the system is completely filled with water with no air bubbles; | ||
## the pump is turned off; | ## the pump is turned off; | ||
# Spin the rotor to 2000rpm | # Spin the rotor to 2000rpm | ||
# Introduce into the system ONE of the following solutions: (a) 100ml of Hydrogen peroxide, 6% solution, ''or'' (b) 100ml of (non-diluted) bleach | # Introduce into the system ONE of the following solutions: (a) 100ml of Hydrogen peroxide, 6% solution, ''or'' (b) 100ml of (non-diluted) bleach | ||
#* Note: Bleach or peroxyde are better than ethanol for cleaning, as cleaning the chamber with ethanol may increase pellet formation at the distal end of the chamber (the manual doesn't explain why, though). | #* Note: Bleach or peroxyde are better than ethanol for cleaning, as cleaning the chamber with ethanol may increase pellet formation at the distal end of the chamber (the manual doesn't explain why, though). | ||
# After the bleach has been pumped in, pump 1L of distilled water. | # After the bleach has been pumped in, pump 1L of distilled water. | ||
# To prevent mold from accumulating, open the lid of the centrifuge to let it dry. The centrifuge should also be turned off at this point. | # To prevent mold from accumulating, open the lid of the centrifuge to let it dry. The centrifuge should also be turned off at this point. | ||
# If the elutriation is not to be used for a long time, it should be taken apart, cleaned and dried. See Hannah in the Bell lab or Francois in the Endy lab to learn how to do this. | |||
Revision as of 13:48, 13 February 2006
Back to: JE-5.0 elutriation system
- Flush through the system 10L of distilled water to remove all traces of buffer and cells.
- Make sure no cells are left at the distal end of the chamber (cells often pellet at that location). If there are still cells, flush through more water with a higher pump rate. If that doesn't work, follow the protocol Cleaning the elutriation chamber.
- Now make sure that:
- the system is completely filled with water with no air bubbles;
- the pump is turned off;
- Spin the rotor to 2000rpm
- Introduce into the system ONE of the following solutions: (a) 100ml of Hydrogen peroxide, 6% solution, or (b) 100ml of (non-diluted) bleach
- Note: Bleach or peroxyde are better than ethanol for cleaning, as cleaning the chamber with ethanol may increase pellet formation at the distal end of the chamber (the manual doesn't explain why, though).
- After the bleach has been pumped in, pump 1L of distilled water.
- To prevent mold from accumulating, open the lid of the centrifuge to let it dry. The centrifuge should also be turned off at this point.
- If the elutriation is not to be used for a long time, it should be taken apart, cleaned and dried. See Hannah in the Bell lab or Francois in the Endy lab to learn how to do this.
Other notes:
- Beckman has tested these products (bleach and hydrogen peroxide) to make sure they don't damage the rotor (and I suppose the elutriation chambers), but they can't guarantee that this procedure is enough to guarantee sterility.
References
- JE-5.0 Elutriation System manual from Beckman-Coulter, 2004
- H.Blitblau, Bell Lab (MIT), 2005
- F. St-Pierre, Endy Lab (MIT), 2005
