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Date of revision and approval
Document reference: WIMR-SWP-OP-WS-506.07
Date of approval: To be approved
Approval authority: Scientific Platforms Manager, Workplace Health and Safety Manager, Advanced Specialist Flow Cytometry
Functional unit: Flow Cytometry
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Enquiries contact: westmead.cytometry@sydney.edu.au
Objective
This document describes the safe operation of the GentleMACS Octo Dissociator located at the Westmead Research Hub Flow Cytometry Core Facility. This document includes starting up the system, basic operation, cleaning, and shutting down the instruments. All personnel require training prior to independent operation of the instruments. OGTR requirements for safe work in a PC2 laboratory apply.
Hours of operation and emergency contacts
The following hours of operation are valid from July 2nd 2018 unless otherwise updated.
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Weekends and public holidays
Training and competency requirements
All new users require instrument training with a relevant training record completed. Initial access is not limited to business hours. Initial usage of the instrument is to be with a trained user.
Instrument booking guidelines
Bookings can be made on PPMS.
List of hazards and risk controls as per risk assessment
Task or scenario | Hazard/s | Associated harm | Existing risk controls | Current risk rating | Additional risk controls? | Residual risk rating? | |||||
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| Instrument operation | Electrocution | Contact with electricity can cause electric shock and burns | Routine instrument maintenance - to ensure instrument is in good condition and cabling is not damaged. Electrical equipment annual testing. Educate users to prevent liquid spill into the instrument. Safety circuit breakers and fuses on instrument to prevent general electrocution due to instrument failure, especially in the presence of liquid. Emergency power off button located in laboratories to disconnects power to the red power points and not the blue uninterruptible power points. | low | |||||||
| Mechanical hazard | Moving parts can cause injury to body parts | Educate users to keep the Trough, Sleeves, and Safety Shield attached when operating. When the instrument’s rotors are rotating, do not touch them or load or remove tubes from the instrument. Always stop or abort a procedure before any adjustments. Warning signs on instrument moving parts. | low | ||||||||
Exposure to bio-hazardous material | Exposure to biohazardous material can cause health issues. | PPE – gloves, gown & enclosed shoes. Biological spill kit - Access to emergency biological spill kit and/or cleaning equipment. Ethanol decontamination of instrument performed during shutdown. Project approval process. | low | ||||||||
Hot surface hazard | The heating foil of the gentleMACS heaters may become hot enough to cause burns if touched. | Educate users not to touch the heating foil, while the instrument is in operation. Warning signs on heating parts | low | ||||||||
Handling hazardous chemicals including Bleach, Decon/Contrad, Ethanol | Hazardous substance exposure | Eye exposure causing eye damage Contact with skin can cause irritation or burn | Safety goggles are provided for researchers and recommended for use in the lab. Emergency showers & eye wash stations available. Chemical spill kit available in shared lab J.2.06. PPE – gloves, gown & enclosed shoes are necessary for working in the laboratory. SDS available to users to ensure awareness of relevant chemical hazards and emergency procedures. | low | Sample handling | Contact with bio-hazardous material | Exposure to bio-hazardous material | Handle samples in biosafety cabinet PPE – gloves, gown & enclosed shoes are necessary for working in the laboratory. Access to emergency biological spill kit and materials to clean up spills. | low |
Procedure
Definitions
In these procedures the following terms have the meaning set out below
- C tubes- Purple-cap C Tubes to get viable single-cell suspensions for cell separation, culture, or analysis.
- M tubes - Orange-cap M Tubes to achieve thorough sample homogenization for subsequent molecular and microbiology analysis.
List of Resources Required
(including personal protective clothing, chemicals and equipment needed)
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- Sample isolation on the GentleMACS® Octo Dissociator with heaters must only occur after the approval of an associated project in PPMS.
Procedure (Step by step instructions or order for undertaking the task)
PPE including gloves, long sleeve gowns, & enclosed shoes must be worn before operating the instrument.
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Samples that are to be isolated are required to be transported to the flow cytometry labs in accordance with PC2 lab requirements in an appropriate container.
Starting up the instrument
- Ensure that the device is plugged in before setting the switch to the ON position (“ ” indicates “ON”, “ ” indicates “OFF”).
- The instrument starts within a few seconds and displays the Main menu.
- The gentleMACS Octo Dissociator with Heaters is operated using the touchscreen display. Items that can be selected are displayed in yellow, while items that cannot
be selected are displayed in white.
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Different folders can be accessed by pressing a folder icon. The arrow indicates scroll function. The Miltenyi & Template folder contains gentleMACS Programs
provided by Miltenyi Biotec. You can also store userdefined programs in the user folder.
Menu tabs
Performing dissociation
Tips and hints for the processing of samples:
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- Attach the tubes upside down to the Sleeves of the free tube positions. The status of the tube positions will change from Free to Selected.
- Refer to the kit for the appropriate program for dissociation.
- To select a dissociation program, press the Folder icon to select the required folder. The list of gentleMACS Programs in the respective folder will be displayed.
- Highlight the desired gentleMACS Program touching the screen or using the scroll buttons.
- Select the required program and press OK to confirm. The program will be assigned to all positions with tubes. The status of the tube positions will change to Ready.
- Press Start to commence the program. The status of the tube positions will change to Running. The progress bar and timer indicate the time remaining.
- On completion of the program, the status of the tube positions will change to Done. The progress bar and timer display the elapsed time since completion of the program.
- Remove the tubes from the tube positions. The status of the tube positions will change to Free.
Cleaning & switching the instrument off
- In case of spillage or contamination, clean the instrument appropriately.
- For decontamination, wipe the instrument with 70% ethanol. Clean the instrument only with cleaning agents that are non-corrosive and not damaging to the plastic
housing of the gentleMACS Octo Dissociator with Heaters. Suitable cleaning agents include household detergents and 70% ethanol. - Power off the instrument
Emergency procedures
Emergency procedure folder can be found here in the sample prep lab:
You can also access all these information via WIMR intranet:
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If there is an electrical hazard and the instrument is plugged into a power point that is not on the uninterruptible power supply (blue power point), pressing the emergency power off button will stop electricity to the instrument. Note that if an electrical device is plugged into the blue power point it will continue to be live. In this event leave the room, ensure a message is placed on the door alerting others and seek assistance.
Additional Information
Instrument failure
In the event of instrument failure, do not attempt to fix the instrument. Report all incidents and instrument failures to the instrument specific emergency contacts listed above.
Associated Documents
This Standard Operating Procedure should be read in conjunction with:
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