Sorting Requirements

  • Pre-Sort Approval
  • Biosafety Table
  • New User Sort Form
  • Sorting Procedures
  • Cell Sorting SOP

The cell sorting room E-386A & B is approved for work at Biosafety Level 1 and 2 (BSL-1 and BSL-2). Room E-377A is additionally approved for Biosafety Level 2 w/ 3 practices. The sorters are housed within class II biosafety cabinets. All cells exposed to or infected with bacterial or viral agents must be approved by EH&S on a case by case basis.

Approval to Sort

The following are required from each user:

  • Creation of a User Account
  • Completion of facility orientation and training.
  • Biological Use Authorization (BUA) from Environmental Health & Safety (EH&S) for samples that are not listed as exempt from the “Biosafety Levels” table in the next Tab.
  • UW Bloodborne Pathogens (BBP) Training documentation (current, within 12 months) for any person working with human cells or bloodborne pathogens.

Create a User Account. 
By filling out the Cell Analysis Facility New User Form, an individual user enables facility staff to create a user account on the CAF server and each of the cytometers.

Facility Orientation & Training
Cell Analysis Facility training will be provided by the CAF Manager or Staff. It will include an overview of the registration system, training on operation of the cell sorter, safety requirements, training on the Biosafety Cell Sorting SOP, and on the site-specific BBP Exposure Control Plan. This training will be documented on the Cell Analysis Facility User Form. The training records will be maintained by the flow facility staff.

EH&S Biological Use Authorization (BUA). 
The National Institutes of Health Office of Biotechnology Activities (NIH OBA) mandates that the Institutional Biosafety Committee (IBC) review and approve all research that involves recombinant DNA. The UW IBC reviews and approves all research involving infectious agents.

PI's are responsible for obtaining BUA's, which apply to all members of their research groups. 

Contact EH&S Research and Biological Safety Office (RBSO) via email at ehsbio@uw.edu , via UW mailbox at 357165, or via fax at 206-221-3068, to obtain a BUA. Documentation of BUA must be in place before users are permitted to sort. Each BUA includes information on cell lines and infectious agents used by each laboratory, their appropriate level of containment, and sites at which they can be used. See the EH&S website for BUA forms and more information.

Bloodborne pathogens (BBP’s)
Staff safety for work involving blood and other potentially infectious materials that may contain Bloodborne pathogens (BBP’s) is mandated by the State of Washington Department of Safety and Health and enforced by the Department of Labor and Industries (WAC-296-823). All researchers and facilities at the UW must comply with these regulations.

Bloodborne Pathogens (BBP) Training Documentation
Employees who have potential for exposure to human blood or other potentially infected materials which includes human cells and cell lines must be enrolled in the UW BBP Program. The Program includes offering of the Hepatitis B vaccine, initial and annual training, and a written site-specific Exposure Control Plan (ECP). The Cell Analysis Facility has an ECP that will be reviewed with users as part of the facility orientation and training.

 

Type of cell/ procedure

Exempt BSL-1

BSL-1

BSL-2

BSL2 with 3 practices
(BSL2+)

Examples of Cells

Wild type cells from murine or other non-human / non-primate species that have NOT been exposed to any microbial agent (e.g., viral, bacterial, fungal, protozoan, parasitic) and have not been genetically modified
or
Cells determined by EH&S and the IBC to be recombinant NIH-exempt BSL-1.

Cells from murine or other non-human / non-primate species that have NOT been exposed to any microbial agent but have been genetically modified using non-viral methods (e.g. cells from transgenic animals or cells treated with nucleic acids).
or
Cells determined by EH&S and the IBC to be approved as non-recombinant BSL-1 or recombinant BSL-1.

Cells from human or non-human primates
or
Cells that have been genetically modified using viral methods
or
Cells exposed to microbial  agents (e.g., viral, bacterial, fungal, protozoan, parasitic)
AND
Have been approved by EH&S and the IBC for BSL-2 containment and sorting

Cells exposed to microbial agents (e.g., viral, bacterial, fungal, protozoan, parasitic) with moderate increased personal hazard.
(e.g. West Nile Virus)
or
Cells that have been genetically modified using viral methods with known oncogenic inserts

AND
Have been approved by EH&S and the IBC for BSL-2+ containment and sorting

BUA update to sort in the Immunology CAF

Not required.

Required.

Required.

Required.

Bloodborne Pathogen (BBP) Training

Not required.

Not required.

Initial and annual renewal required for work with human cells and other cells exposed to bloodborne pathogens.

Initial and annual renewal required for work with human cells and other cells exposed to bloodborne pathogens.

Sort Sign-up

Required. Include name, phone number, sample description.

Required. Include name, phone number, sample description and a clear BSL-1 notation.

Required. Include name, phone number, sample description and a clear BSL-2 notation.

Required. Include name, phone number, sample description and a clear BSL-2+ notation.

Sample Transport

No requirement

Leak-proof secondary container.

Leak-proof secondary container.

Leak-proof secondary container.

Room Restriction

None.

None.

YES – door to E-386A or B must be closed during sort and the BSL-2 Biohazard sign posted on the outside.

YES – door to E-377A must be closed during sort and the BSL-2 with BSL- 3 practices Biohazard sign posted on the outside.

Personal Protective Equipment (PPE)

Closed toe shoes.  When manipulating samples (i.e. loading/unloading samples), gloves, lab coat, and face protection are recommended.

Closed toe shoes. When manipulating samples (i.e. loading/unloading samples), gloves, lab coat, and face protection are strongly recommended.

Spills: Lab coat, nitrile gloves, goggles/faceshield required.

Closed toe shoes, lab coat and gloves required at all times. When manipulating samples (i.e. loading/unloading samples), face protection is also required.

Spills: Lab coat, nitrile gloves, goggles/faceshield required.

Closed toe shoes, shoe covers, disposable rear opening gown, double gloves and face protection is required at all times.

 

Spills: Lab coat, nitrile gloves, goggles/faceshield required.

Waste

Contaminated materials must be decontaminated prior to disposal.

Gloves and other waste must be disposed of as biohazardous. See CAF SOP.

Gloves and other waste must be disposed of as biohazardous. See CAF SOP.

Gloves and other waste must be disposed of as biohazardous. See CAF SOP.

 

Please use the New User Sorting Approval Form to provide the information the faciltiy will need to determine if your samples will require an updated Biological Use Authorization from EH&S.

Pre-Sort  Procedures

After the pre-sort approval is complete, the following procedures must be followed by facility users prior to sorting.

1. Sign-up:  Each user is responsible for signing up using the online scheduling system   (https://depts.washington.edu/flowlab/Schedule/CalciumView.cgi/inst_signup).  Users sign up for sorting time in 15 minute increments, with an extra 15 minutes included at the end of the sort for decontamination and cleaning. 

  1. Sign-up information MUST include: user name, sample description, and a contact phone number.
  2. Each entry is approved after the Facility Staff has looked at the request and made sure that there will be appropriate staff and all the required information is provided.
  3. All BSL-2 sorts must include the BSL-2 notation as well as additional notation further clarifying the sample description (e.g.- ES cells, patient samples, transfected cells, infected cells etc). 

Cell Analysis Facility staff is available for presort consultation.


You MUST contact CAF staff at 543-7006 prior to scheduling a same day sort to ensure you can be accommodated.

All sorts scheduled are forwarded to the staff via e-mail for approval prior to appearing on the calendar. To expedite your request to reserve time provide detailed information regarding your sort as listed above.

Once you have submitted a request that time slot is secured and will appear on the calendar after the staff verifies that you can be accommodated.

Please note the following:

If you cancel less than 24 hours in advance, you will be charged for 1 hour of set-up time.

No shows will be charged for the full amount of time they have reserved.

2. Review the following documents:

Before you Sort

Aria Sample Prep this document details many common questions related to sorting, please read!

Please review the Manual of Standard Operating Procedures for sorting in the SLU Cell Analysis Facility.

Contact Michele Black or ehsbio@uw.edu if you have any questions.

 

 

 

 

 

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