Protocol Citation: tdoerks 2022. NARMS-KS-1-Autoclave Use. protocols.io https://protocols.io/view/narms-ks-1-autoclave-use-ci4mugu6
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: November 09, 2022
Last Modified: November 19, 2022
Protocol Integer ID: 72557
Abstract
Instructions on how to use autoclave in Mosier P209.
Waste
Waste
Make sure waste to be autoclaved is in a biohazard bag (no more than 3/4 full) and place in autoclavable container.
Wrap 2 Steam Migrating Integrator Strips together with autoclave tape and a piece of string and place in the middle of the bag of waste to be autoclaved with the piece of string hanging out of the bag so that the strips are easily retrieved after autoclave cycle is finished
Tape string with autoclave tape to an AccuFast Biological Indicator (BI) and place in the middle of the bag of waste to be autoclaved with the piece of string hanging out of the bag so that the BI is easily retrieved after autoclave cycle is finished.
Add 1 liter of water to the bag and loosely close with autoclave tape leaving about a 2-finger space opening and load from door in room P209.
Close the door with foot peddle and select Cycle 4 from the cycle menu and select 'Start Cycle.'
Cycle Parameters:
Cycle Type - Gravity
Purge - 1:00
Sterilization Time - 60:00
Sterilization Temp - 121.0°C
Overdrive - 1.5°C
Overtemp - 6.0°C
Undertemp - 1.0°C
Undertemp - Restart
Print Interval - 2 minutes
Vacuum Dry - 10.0 inHG
Dry Time - 05:00
When load is finished, open door with caution as steam can be released from the doorway and using appropriate PPE (temperature resistant gloves, lab coat, and goggles) unload the tray containing the bag.
Observe autoclave tape color and record results (color change - yes or no).
Remove and read the steam strips and record results.
Blue in the 'Pass' window indicates 'Pass' otherwise it is considering 'Fail.'
If one steam strip fails and one strip passes, record as 'Pass.'
Remove BI and crack open glass ampule using the indicated slot in the incubator (L202) and incubate at 60°C for 10 hours.
Blue color indicates 'Pass' while yellow color indicates 'Fail.'
In the case where BI and the steam strip results conflict, defer to the BI results as they are considered more accurate and consistent.
Example:
BI=Pass, Steam Strip=Fail, then consider the load as 'Pass'
BI=Fail, Steam Strip=Pass, then consider the load as 'Fail'
Record all results on NARMS-KS-F-1A Autoclave Log
If load has passed sterilization then it can be placed in a black trash bag and taken out to the trash. If load has not passed then it needs to be reautoclaved until it passes sterilization. Please see section on trouble shooting should this occur.
Media
Media
Place all media in autoclavable trays and make sure to not overload autoclave with media like stacking trays on top of each other (only one layer thick) or putting media bottles too close to each other (make sure they are not touching with at least a finger width or two apart)
Place autoclave tape on each bottle or rack of media to be autoclaved along with 2 Steam Migrating Integrator Strips wrapped together with autoclave tape and a piece of string and place in the middle of one of the trays to be autoclaved with the piece of string hanging out of the tray so that the strips are easily retrieved after autoclave cycle is finished.
Tape string with autoclave tape to an AccuFast Biological Indicator (BI) and place in the middle of one of the trays containing media with the piece of string hanging out of the tray so that the BI is easily retrieved after autoclave cycle is finished.
Close the door with foot peddle and select Cycle 5 from the cycle menu and select 'Start Cycle.'
Cycle Parameters:
Cycle Type - Liquid
Purge - 1:00
Sterilization Time - 15:00
Sterilization Temp - 121.0°C
Overdrive - 1.5°C
Overtemp - 6.0°C
Undertemp - 1.0°C
Undertemp - Restart
Print Interval - 2 minutes
When load is finished, open door with caution as steam can be released from the doorway and using appropriate PPE (temperature resistant gloves, lab coat, and goggles) unload the trays containing the media and let cool before handling.
Observe autoclave tape color and record results (color change - yes or no).
Remove and read the steam strips and record results.
Blue in the 'Pass' window indicates 'Pass' otherwise it is considering 'Fail.'
If one steam strip fails and one strip passes, record as 'Pass.'
Remove BI and crack open glass ampule using the indicated slot in the incubator (L202) and incubate at 60°C for 10 hours.
Blue color indicates 'Pass' while yellow color indicates 'Fail.'
In the case where BI and the steam strip results conflict, defer to the BI results as they are considered more accurate and consistent.
Example:
BI=Pass, Steam Strip=Fail, then consider the load as 'Pass'
BI=Fail, Steam Strip=Pass, then consider the load as 'Fail'
Record all results on NARMS-KS-F-1A Autoclave Log
If load has passed sterilization then media is acceptable for use. If load has not passed then the media needs to be reautoclaved until it passes sterilization. Please see section on trouble shooting should this occur.
Glassware and Instruments
Glassware and Instruments
Place all glassware and/or instruments in autoclavable trays and make sure to not overload autoclave with items like stacking trays on top of each other (only one layer thick) or putting items too close to each other.
Place autoclave tape on each item or autoclave tray to be autoclaved along with 2 Steam Migrating Integrator Strips wrapped together with autoclave tape and a piece of string and place in the middle of one of the trays to be autoclaved with the piece of string hanging out of the tray so that the strips are easily retrieved after autoclave cycle is finished.
Tape string with autoclave tape to an AccuFast Biological Indicator (BI) and place in the middle of one of the trays containing glassware and/or instruments with the piece of string hanging out of the tray so that the BI is easily retrieved after autoclave cycle is finished.
Close the door with foot peddle and select Cycle 3 (use Cycle 4 for large loads) from the cycle menu and select 'Start Cycle.'
Cycle Parameters:
Cycle Type - Gravity
Purge - 1:00
Sterilization Time - 30:00
Sterilization Temp - 121.3°C
Overdrive - 1.5°C
Overtemp - 6.0°C
Undertemp - 1.0°C
Undertemp - Restart
Print Interval - 2 minutes
Vacuum Dry - 10.0 inHG
Dry Time - 18:00
When load is finished, open door with caution as steam can be released from the doorway and using appropriate PPE (temperature resistant gloves, lab coat, and goggles) unload the trays and let cool before handling.
Observe autoclave tape color and record results (color change - yes or no).
Remove and read the steam strips and record results.
Blue in the 'Pass' window indicates 'Pass' otherwise it is considering 'Fail.'
If one steam strip fails and one strip passes, record as 'Pass.'
Remove BI and crack open glass ampule using the indicated slot in the incubator (L202) and incubate at 60°C for 10 hours.
Blue color indicates 'Pass' while yellow color indicates 'Fail.'
In the case where BI and the steam strip results conflict, defer to the BI results as they are considered more accurate and consistent.
Example:
BI=Pass, Steam Strip=Fail, then consider the load as 'Pass'
BI=Fail, Steam Strip=Pass, then consider the load as 'Fail'
Record all results on NARMS-KS-F-1A Autoclave Log
If load has passed sterilization then media is acceptable for use. If load has not passed then the media needs to be reautoclaved until it passes sterilization. Please see section on trouble shooting should this occur.
Troubleshooting
Troubleshooting
Load does not pass sterilization
Confirm correct load was selected and ran with no errors
Reduce the amount of waste in the bag (separate into two bags?)
Add more or less water into the bag and between all 'steam barriers (smaller bags inside larger bags etc) in order that steam reaches all waste in the bag
Screen says 'CLOSE THE DOOR BEFORE STARTING CYCLE'
If door in P209 is closed, then push up slightly on the door in P209A (other side of autoclave) and attempt to start the cycle
If load stops in the middle of cycle with alarm sounding and says 'TOO LONG TO SEAL DOOR'
Press 'Service Help', then 'Clear Alarm', followed by 'Abort', then 'Abort' again and 'Status Print' and wait for instruemtn to return to original screen.
Check door in P209A fully closed by pushing up slightly and that the seal around the door in P209 is pushed in and has not obstructed the door and restart the load.