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Information Operating
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SetupAssembleClosedSystemOperatingManual.SetupAssembleClosedSystem HistoryHide minor edits - Show changes to output Changed line 70 from:
#Wrap all culture vessel to:
#Wrap all culture vessel effluent tube openings in aluminum foil Changed line 72 from:
**Double check that all open ends of the tubing system, to:
**Double check that all open ends of the tubing system, effluent ports, and air filters are properly wrapped in aluminum foil Added line 80:
#As soon as the autoclave finishes remove the foil from '''only''' the air filters. This will allow them to dry. Only dry filters will pass air. Changed lines 65-68 from:
# #Connect an air filter to the other inport port on the culture vessel #Repeat for to:
# For each culture tube ##Connect one of the Female Luer ends from the culture tube to one of the male ends on the tubing manifold. ##Connect an air filter to the other Female Luer port on the culture vessel **After this step, the culture vessels should only have their effluent ports open Changed lines 52-54 from:
##Wash with soap and water ##Rinse culture tube with ethanol to:
##Wash with soap and water using a tube brush Changed lines 54-55 from:
#Clean magnetic stir bar to:
#Clean magnetic stir bar with soap and water Changed line 56 from:
#Place stir bar into culture tube with a little bit of DI to:
#Place stir bar into culture tube with a little bit of DI water (whatever is left from rinsing is okay) Changed lines 58-61 from:
->[-''''' #Rinse out input ports with ethanol #Securely place plug to:
->[-'''''Stopper'''''-] #Rinse out input ports with ethanol from a squeeze bottle #Rinse out export port with ethanol from a squeeze bottle #Securely place Stopper into culture tube Changed line 44 from:
#Fasten to:
#Fasten a Male Luer lock connector to tubing created in step 1 Changed line 46 from:
#Attach to:
#Attach the female end of the Teflon air filter to other end of the tubing Changed lines 17-18 from:
**( to:
**(1) 1" tube **(1) 3" tube Changed line 20 from:
**(1) 5' to:
**(1) 5' tube Added lines 22-26:
## Attach two 6-way barb manifolds end-to-end with the 1" section of tube. ## On the two other ends attach the 3" and 5' tubes. ## On the remaining 8 ends of the barb manifolds attach the 8 2' tubes. ## On the 8 2' pieces of tubing attach male Luer-barb connectors. ## on the 2 remaining pieces, connect female Luer-barb connectors. Changed line 42 from:
#Cut tubing ( to:
#Cut tubing (1/8" ID x 1/4" OD) Changed line 16 from:
# Cut Tubing ( to:
# Cut Tubing (1/16" ID 1/8" OD silicone). Note: you may adjust the length of tubing as needed for your configuration. Changed line 15 from:
->[-'''''If preparing for the first time to:
->[-'''''If preparing for the first time.'''''-] Added lines 22-26:
->[-'''''If reusing tubing.'''''-] # Inspect the tubing. ## Check the tubing for holes and leaks. ## Check the tubing for fatigue cause by the pinch valves. Look for dents in the tubing, if it looks bad enough or tears when stretched, replace the tubing. Changed lines 30-32 from:
## Rinse with DI to:
## Rinse with DI water. ### Note: It's important to thoroughly rinse all tubing. Any left over media will burn in the autoclave and may discolor the media used to blank the system. Changed line 20 from:
# Assemble to:
# Assemble tubing manifold Changed line 22 from:
# Rinse to:
# Rinse tubing system Added line 24:
### using a squeeze bottle flush ethanol into each opening of the 10 way tubing manifold. Changed line 10 from:
[-'''Six Way to:
[-'''Six Way Manifold'''-]\\\ Deleted line 36:
Changed line 67 from:
**Set the temperature to 121 deg to:
**Set the temperature to 121 °C? Added lines 6-7:
[-'''Male Luer Lock'''-]\\\ %width=150px%Attach:LuerLockMale2.jpeg Deleted lines 8-9:
[-'''Male Luer Lock'''-]\\\ Added line 37:
#NEED TO EXPLAIN HOW TO CREATE STOPPER WITH NEEDLES IN IT Changed line 1 from:
[[OperatingManual/ to:
[[OperatingManual/SetupAttachOpenEnds | Next]] Changed line 70 from:
[[OperatingManual/ to:
[[OperatingManual/SetupAttachOpenEnds | Next]] Changed line 1 from:
[[ to:
[[OperatingManual/SetUpAttachOpenEnds | Next]] Changed line 70 from:
[[ to:
[[OperatingManual/SetUpAttachOpenEnds | Next]] Changed line 6 from:
[-Female Luer Lock-]\\\ to:
[-'''Female Luer Lock'''-]\\\ Changed line 8 from:
[-Male Luer Lock-]\\\ to:
[-'''Male Luer Lock'''-]\\\ Changed line 10 from:
[-Six Way Connector-]\\\ to:
[-'''Six Way Connector'''-]\\\ Added line 1:
[[https://students.washington.edu/cnt/pmwiki/pmwiki.php?n=OperatingManual.SetupAttachOpenEnds | Next]] Changed line 70 from:
[[https://students.washington.edu/cnt/pmwiki/pmwiki.php?n=OperatingManual.SetupAttachOpenEnds | to:
[[https://students.washington.edu/cnt/pmwiki/pmwiki.php?n=OperatingManual.SetupAttachOpenEnds | Next]] Changed lines 67-69 from:
**Setting is a preset called "Packs" on some to:
**Setting is a preset called "Packs" on some autoclaves [[https://students.washington.edu/cnt/pmwiki/pmwiki.php?n=OperatingManual.SetupAttachOpenEnds | Continue to next section]] Changed line 6 from:
%width=150px% to:
%width=150px%Attach:LuerLockMale2.jpeg Changed line 8 from:
%width=150px% to:
%width=150px%Attach:6WayConnector.jpeg Changed lines 10-11 from:
% to:
%width=150px%[[https://students.washington.edu/cnt/pmwiki/pub/uploads/OperatingManual/SetupAssembleClosedSystem/AssembledTubing.jpeg | https://students.washington.edu/cnt/pmwiki/pub/uploads/OperatingManual/SetupAssembleClosedSystem/AssembledTubing.jpeg"Click to enlarge"]] [-Assembled Tubing-]\\\ %width=150px%[[https://students.washington.edu/cnt/pmwiki/pub/uploads/OperatingManual/SetupAssembleClosedSystem/InputConnection.jpeg | https://students.washington.edu/cnt/pmwiki/pub/uploads/OperatingManual/SetupAssembleClosedSystem/InputConnection.jpeg"Click to enlarge"]] [-Input port connection-] Changed lines 10-11 from:
%thumb width=150px% [[https://students.washington.edu/cnt/pmwiki/pub/uploads/OperatingManual/SetupAssembleClosedSystem/AssembledTubing.jpeg | https://students.washington.edu/cnt/pmwiki/pub/uploads/OperatingManual/SetupAssembleClosedSystem/AssembledTubing.jpeg"Click to enlarge"]] [-Assembled Tubing-] to:
%thumb width=150px% [[https://students.washington.edu/cnt/pmwiki/pub/uploads/OperatingManual/SetupAssembleClosedSystem/AssembledTubing.jpeg | https://students.washington.edu/cnt/pmwiki/pub/uploads/OperatingManual/SetupAssembleClosedSystem/AssembledTubing.jpeg"Click to enlarge"]] [-Assembled Tubing-]\\\ Attach:InputConnection.jpeg Changed line 4 from:
%width=150px%Attach:LuerLockFem1. to:
%width=150px%Attach:LuerLockFem1.jpeg Changed line 6 from:
%width=150px% Attach:LuerLockMale2. to:
%width=150px% Attach:LuerLockMale2.jpeg Changed line 8 from:
%width=150px% Attach:6WayConnector. to:
%width=150px% Attach:6WayConnector.jpeg Changed line 1 from:
!!!NEED: Picture of air filters, diagram of input and export holes, and completed closed system to:
!!!NEED: Picture of air filters, diagram of input and export holes on plug, and completed closed system Added lines 1-2:
!!!NEED: Picture of air filters, diagram of input and export holes, and completed closed system Changed line 16 from:
# Assemble tubes to:
# Assemble tubes together in an "octopus" system Changed line 18 from:
# Rinse to:
# Rinse octopus tubing system Changed line 50 from:
#Connect a Luer lock end of a 2' tubing section to an inport port on the culture vessel to:
#Connect a Luer lock end of a 2' tubing section of the octopus tubing system to an inport port on the culture vessel Changed line 61 from:
#Place closed to:
#Place closed tubing-culture vessel system into an autoclave Changed line 63 from:
#Run autoclave for 30 to:
#Run autoclave for 30 minutes Changed line 44 from:
#Rinse out to:
#Rinse out input ports with ethanol Deleted lines 45-47:
->[-'''''Assembly'''''-] #Place stir bar into culture tube Changed lines 33-37 from:
#Clean culture tube with **Rinse culture tube with ethanol ** to:
#Clean culture tube ##Wash with soap and water ***Make sure to clean all biofilms off of culture tube walls ##Rinse culture tube with ethanol ##Rinse culture tube with DI water Changed lines 39-40 from:
to:
##Rinse with ethanol ##Rinse with DI water Changed line 34 from:
**Take care to clear all biofilms off of culture to:
**Take care to clear all biofilms off of culture tube walls Changed lines 36-39 from:
# #Place stir bar into culture to:
**Rinse culture tube with ethanol **Rinse culture tube with DI water #Clean magnetic stir bar **Rinse with ethanol #Place stir bar into culture tube with a little bit of DI water Changed line 16 from:
# to:
# Assemble tubes together Changed lines 1-2 from:
%center%%block rframe width=150px% %width=150px%Attach:LuerLockFem1.jpeg\\ to:
%center%%block rframe width=150px% [+{+'''Parts'''+}+]\\ %width=150px%Attach:LuerLockFem1.jpeg\\ Added line 9:
Changed line 7 from:
%width=150px% to:
%thumb width=150px% [[https://students.washington.edu/cnt/pmwiki/pub/uploads/OperatingManual/SetupAssembleClosedSystem/AssembledTubing.jpeg | https://students.washington.edu/cnt/pmwiki/pub/uploads/OperatingManual/SetupAssembleClosedSystem/AssembledTubing.jpeg"Click to enlarge"]] [-Assembled Tubing-] Changed line 1 from:
%block rframe width=150px% %width=150px%Attach:LuerLockFem1.jpeg\\ to:
%center%%block rframe width=150px% %width=150px%Attach:LuerLockFem1.jpeg\\ Changed line 2 from:
Female Luer Lock\\\ to:
[-Female Luer Lock-]\\\ Changed line 4 from:
Male Luer Lock\\\ to:
[-Male Luer Lock-]\\\ Changed lines 6-7 from:
Six Way Connector\\\ %width=150px% Attach:AssembledTubing.jpeg Assembled to:
[-Six Way Connector-]\\\ %width=150px% Attach:AssembledTubing.jpeg [-Assembled Tubing-] Added lines 1-7:
%block rframe width=150px% %width=150px%Attach:LuerLockFem1.jpeg\\ Female Luer Lock\\\ %width=150px% Attach:LuerLockMale2.jpeg\\ Male Luer Lock\\\ %width=150px% Attach:6WayConnector.jpeg\\ Six Way Connector\\\ %width=150px% Attach:AssembledTubing.jpeg Assembled Tubing Changed lines 15-20 from:
**See images %rframe width=150px% Attach:LuerLockFem1.jpeg | Female Luer Lock %rframe width=150px% Attach:LuerLockMale2.jpeg | Male Luer Lock %rframe width=165px% Attach:6WayConnector.jpeg | Six Way Connector %rframe width=180px% Attach:AssembledTubing.jpeg | Assembled Tubing to:
**See images to the left: Changed lines 25-28 from:
**See Images %rframe width=150px% Attach to:
**See Images to the left: Changed lines 10-14 from:
% % % %cframe \\ to:
%rframe width=150px% Attach:LuerLockFem1.jpeg | Female Luer Lock %rframe width=150px% Attach:LuerLockMale2.jpeg | Male Luer Lock %rframe width=165px% Attach:6WayConnector.jpeg | Six Way Connector %rframe width=180px% Attach:AssembledTubing.jpeg | Assembled Tubing Changed lines 10-13 from:
% % % % to:
%cframe width=150px% Attach:LuerLockFem1.jpeg | Female Luer Lock %cframe width=150px% Attach:LuerLockMale2.jpeg | Male Luer Lock %cframe width=165px% Attach:6WayConnector.jpeg | Six Way Connector %cframe width=180px% Attach:AssembledTubing.jpeg | Assembled Tubing Changed line 13 from:
%rframe width to:
%rframe width=300px% Attach:AssembledTubing.jpeg | Assembled Tubing Changed line 12 from:
%rframe width=165px% Attach: to:
%rframe width=165px% Attach:6WayConnector.jpeg | Six Way Connector Changed line 14 from:
to:
\\ Changed line 18 from:
to:
\\ Changed line 11 from:
%rframe width=150px% Attach: to:
%rframe width=150px% Attach:LuerLockMale2.jpeg | Male Luer Lock Changed line 13 from:
%rframe width = to:
%rframe width =300px% Attach:AssembledTubing.jpeg | Assembled Tubing Changed lines 8-14 from:
**See to:
**See images bellow: %rframe width=150px% Attach:LuerLockFem1.jpeg | Female Luer Lock %rframe width=150px% Attach:LuerLockMale1.jpeg | Male Luer Lock %rframe width=165px% Attach:5WayConnector.jpeg | Five Way Connector %rframe width =150px% Attach:AssembledTubing.jpeg | Assembled Tubing Changed lines 23-25 from:
#Fasten Luer lock **See %width=50px%Attach:LuerLockFem1.jpeg to:
#Fasten Luer lock connectors to tubing created in step 1 **See Images Bellow %rframe width=150px% Attach:LuerLockFem1.jpeg | Female Luer Lock Changed lines 64-67 from:
**Setting is a preset called "Packs" on some http://pmichaud.com/img/misc/pc.jpg"Paper clips" | [- %newwin% [[ Wikipedia:Paper_clips | Paper clips ]] are ''fun'' to work with. -] to:
**Setting is a preset called "Packs" on some autoclaves Changed line 19 from:
to:
%width=50px%Attach:LuerLockFem1.jpeg Changed line 59 from:
Wikipedia:Paper_clips | Paper clips ]] are ''fun'' to work with. -] to:
Wikipedia:Paper_clips | Paper clips ]] are ''fun'' to work with. -] Changed lines 56-59 from:
**Setting is a preset called "Packs" on some to:
**Setting is a preset called "Packs" on some autoclaves http://pmichaud.com/img/misc/pc.jpg"Paper clips" | [- %newwin% [[ Wikipedia:Paper_clips | Paper clips ]] are ''fun'' to work with. -] Changed line 19 from:
**"Attach:LuerLockFem1. to:
**"Attach:LuerLockFem1.jpeg" Changed line 19 from:
"Attach: to:
**"Attach:LuerLockFem1.JPG" Changed lines 53-54 from:
**Set the temperature to #Run autoclave for 30? to:
**Set the temperature to 121º C? #Run autoclave for 30? minutes **Setting is a preset called "Packs" on some autoclaves Changed lines 52-54 from:
#Place closed system into an autoclave with to:
#Place closed system into an autoclave with **Set the temperature to 125º C? #Run autoclave for 30? minutes Changed lines 49-52 from:
->'''NOTE:''' All aluminum foil wraps should be tight enough so that no liquid is able to enter the now enclosed tubing-culture vessel to:
->'''NOTE:''' All aluminum foil wraps should be tight enough so that no liquid is able to enter the now enclosed tubing-culture vessel system [+'''@@AUTOCLAVE CLOSED SYSTEM@@'''+] #Place closed system into an autoclave with a set temperature of 125º C? for 30? minutes Changed lines 15-19 from:
# to:
#Cut tubing (2cm diameter?) **(8) 6" tube #Fasten Luer lock connector to tubing created in step 1 **See diagram bellow (not implemented yet) #Attach Teflon air filter to connecter side of tubing Changed lines 1-2 from:
'''@@TUBING PREPARATION@@''' to:
[+'''@@TUBING PREPARATION@@'''+] ->[-'''''If preparing for the first time, do steps 1-2 else skip to 3'''''-] Changed lines 13-14 from:
'''@@ to:
[+'''@@AIR FILTER PREPARATION@@'''+] ->[-'''''Execute steps bellow only if no air filters are created or need replacing'''''-] # [+'''@@CULTURE VESSEL PREPARATION@@'''+] ->[-'''''Body'''''-] Changed line 27 from:
->'''''Plug''''' to:
->[-'''''Plug'''''-] Changed line 31 from:
->'''''Assembly''''' to:
->[-'''''Assembly'''''-] Changed line 36 from:
'''@@CLOSED SYSTEM ASSEMBLY@@''' to:
[+'''@@CLOSED SYSTEM ASSEMBLY@@'''+] Changed lines 14-15 from:
->''''' '''Body to:
->'''''Body''''' Changed lines 22-23 from:
''' to:
#Repeat steps 1-5 for the 7 remaning culture tubes ->'''''Plug''''' Changed lines 26-27 from:
''' to:
#Repeat steps 1-2 for the 7 remaning culture tubes ->'''''Assembly''''' Changed lines 30-31 from:
to:
#Repeat steps 1-2 for the 7 remaning culture tubes Changed line 41 from:
->'''NOTE:''' All aluminum foil wraps should be tight enough so that no liquid is able to enter the now enclosed tubing-culture vessel system to:
->'''NOTE:''' All aluminum foil wraps should be tight enough so that no liquid is able to enter the now enclosed tubing-culture vessel system Changed line 16 from:
#Clean culture to:
#Clean culture tube with water and soap Changed lines 19-20 from:
#Rinse culture #Rinse culture to:
#Rinse culture tube with ethanol #Rinse culture tube with DI water Changed line 22 from:
#Place stir bar into culture to:
#Place stir bar into culture tube Changed lines 27-29 from:
#Place stir bar into culture to:
#Place stir bar into culture tube #Securely place plug into culture tube Changed lines 31-39 from:
#Connect Luer lock to:
#Connect a Luer lock end of a 2' tubing section to an inport port on the culture vessel #Connect an air filter to the other inport port on the culture vessel #Repeat for the 7 remaining culture vessels **After this step, the culture vessels should only have their export ports open to the environment #Wrap all open ends of the tubing system in aluminum foil #Wrap all culture vessel export ports in aluminum foil #Wrap the air filter in aluminum foil **Double check that all open ends of the tubing system, export ports, and air filters are properly wrapped in aluminum foil ->'''NOTE:''' All aluminum foil wraps should be tight enough so that no liquid is able to enter the now enclosed tubing-culture vessel system Changed line 15 from:
''' to:
'''Body''' Changed lines 23-24 from:
''' to:
'''Plug''' #Rinse out inport ports with ethanol Changed line 26 from:
''' to:
'''Assembly''' Added line 31:
#Connect Luer lock ends of the 2' tubing sections to an inport port on the culture vessel Changed lines 15-16 from:
# to:
'''BODY''' #Clean culture vessel with water and soap Added lines 19-30:
#Rinse culture vessel with ethanol #Rinse culture vessel with DI water #Rinse stir bar with ethanol #Place stir bar into culture vessel '''PLUG'' #Rinse out input ports with ethanol #Rinse out export port with ethanol '''ASSEMBLY''' #Place stir bar into culture vessel #Securely place plug into culture vessel '''@@CLOSED SYSTEM ASSEMBLY@@''' Changed lines 15-17 from:
# to:
#Rinse culture vessel with water and soap **Take care to clear all biofilms off of culture vessel ***It's okay for stir bar to be in or out of the culture vessel Deleted lines 0-8:
*50mm? diameter tubing *2cm? diameter tubing *(16) ?gage syringe *(8) Foam stopper *(8) Glass capillary *(8) Teflon air filter Added line 2:
->'''''If preparing for the first time, do steps 1-2 else skip to 3''''' Added lines 1-23:
'''@@MATERIALS@@''' *50mm? diameter tubing *2cm? diameter tubing *(16) ?gage syringe *(8) Foam stopper *(8) Glass capillary *(8) Teflon air filter '''@@TUBING PREPARATION@@''' # Cut Tubing (50mm diameter?) **(2) 3" tube **(4) 2' tube **(1) 5' long tube # Fasten connectors to tubing **See diagram bellow (not implemented yet) # Rinse tubing ## Rinse with ethanol ## Rinse with DI water '''@@CULTURE VESSEL PREPARATION@@''' ->'''''Apply to all culture vessels''''' # |