Policy Acknowledgement- PLEASE READ!

Penn Cytomics & Cell Sorting SRL Policies 

Please review all policies. Signature REQUIRED at bottom for all SRL users.

Access: All instruments located within the centralized laboratory (206/231/297 John Morgan Building) are available 24 hours/day, 365 days/year. Rooms housing these instruments are unlocked Monday-Friday, 8am-6pm. For after-hours access to any of these 3 rooms, you must:

  1. Have a valid PennCard for door access
  2. Complete the required training module for the instrument you are using
  3. Email William.murphy@pennmedicine.upenn.edu to request after-hours access

All Penn Cytomics satellites are accessible Monday-Friday from 9am-5pm. Satellite facilities are only available after hours for users within that building/department, or for users with authorization. Because of long standing support from the Abramson Cancer Center (ACC) project grant, highest priority is given to ACC members with peer-reviewed funded projects, followed by ACC members with other funded projects and non-ACC members with peer-reviewed funded projects, non-ACC members with funded projects, and finally users from outside the institution. There is no discount to ACC members vs. non-members.

Biosafety: Flow cytometric analysis of human and other potentially biohazardous samples is permitted at all of our locations, so long as appropriate precautions are taken. All lab materials must be disposed of correctly, even if they are not contaminated. Each satellite facility has dedicated receptacles for gloves and sharps, which should be provided by that department; FACS tubes and pipette tips are considered sharps. Cell sorting of biohazardous material is only allowed on certain instruments, pending staff approval of a designated biosafety form. Sorting of BSL-2+ materials is only permitted on a dedicated instrument in a biohazardous suite (Aria D) and may only be performed by appropriately trained staff equipped with a PAPR. (Note: The SRL does not support radioactive samples/materials for analyzers and sorting).

Lab Coats/PPE: As mandated by the University of Pennsylvania Environmental Health and Radiation Safety (EHRS) office, lab coats must be worn at all times while using any Penn Cytomics instruments. Users are responsible for providing their own lab coats, gloves, and any other relevant PPE. Eating/drinking in lab areas is strictly prohibited.

Cytometer User Etiquette: 

All users should:

⇒ Fill the sheath and empty the waste container before running their experiments.

⇒ Clean the cytometer after their session.*

⇒ Make sure the cytometer is in STANDBY mode before leaving.

⇒ Take their tubes, tube racks, and waste/trash back with them.

⇒ Shut the cytometer off if they are the last scheduled user of the day.

*Analyzer cleaning steps
In high speed, run the following tubes:

  1. 2% Contrad for 30 seconds with the arm to the side
  2. 2% Contrad for 5 minutes with the arm under
  3. Water for 30 seconds with the arm to the side
  4. Water for 2 minutes with the arm under
  5. FACSClean bleach for 30 seconds with the arm to the side
  6. FACSClean bleach for 5 minutes with the arm under
  7. Water for 30 seconds with the arm to the side
  8. Water for 2 minutes with the arm under

Note: Always run FACSClean bleach when running human samples

For sorter cleaning steps, please refer to the Sorter Training and Resource Guide.

FACS Tubes & Filters for Sample Acquisition: Samples must be in Corning Falcon brand 5mL round-bottom polystyrene (Cat. 352054) or polypropylene tubes (Cat. 352063). Samples must be freshly filtered (at least less than 2 hours) with a 35μm filter BEFORE loading on the cytometer. Beads do not need to be filtered. Falcon Cell-Strainer Cap tubes (Cat. 352235) are recommended for sample filtering. Other brands of tubes may not fit the cytometers appropriately. The use of tube inserts (e.g., cluster tubes, bullet tubes) is not allowed on any Penn Cytomics instruments, as they lead to damage of the BAL seals.

Fisher Polystyrene Tubes (All Analyzers and Sorters, except Influx)

Fisher Polypropylene Tubes (All Sorters, preferred tube for sticky cells)

Instrument Usage via PennCard: Entrée is our usage and data backup software and identifies when a session is taking place. Entrée computer nodes are situated at each instrument workstation. Trained users must scan their PennCard* on the node’s card reader and select a valid funding source to unlock/access instrumentation. NEVER USE YOUR PENNCARD TO LOG IN ANOTHER PERSON, especially if this person hasn’t completed training through our core facility. Failure to comply will result in YOUR instrument privileges being revoked. (See SRL Policy Violations below) If the previous user forgot to log out of Entrée at any instrument workstation, do NOT use their session. Please log them out and log in with your PennCard by tapping it on the card reader. Failure to do so will result in your data going to the previous user, and you will also be billed as a no-show. Please leave an Entrée message at the end of your session to notify the core of any log in/log out issues.

*All PennCards need to be Entrée-registered before gaining access to any Penn Cytomics instruments. This can be done by scanning your PennCard on our registration card reader in 297 JMB. If you lost your badge/card expired and received a new card, you will need to re-register your new card.

Billing: A billing session is initiated the instant a user logs into an instrument using their PennCard, to the moment the user logs out of the cytometer’s acquisition software, regardless of the reason for logging in (i.e. data export). The SRL usage and billing software, termed Entrée, tracks instrument usage to the second, and generates a billable line item to be invoiced monthly. Users are able to select an appropriate funding source to bill when they initiate a session. In order to encourage maximum usage, and to prevent overestimation of time needed on an instrument, SRL policy is to bill a minimum of 75% of scheduled time for underused sessions, and to bill for actual usage for sessions lasting longer than the reserved time. For example, a user who reserves 4 hours of instrument time and uses 1.5 hours will be charged for 3 hours of usage. Users who fail to use reserved instrument time (no-shows) likewise trigger a minimum charge of 75% of scheduled time. Principal Investigators and Departmental BAs will receive a separate monthly invoice notifying them of these charges. See Scheduling & Cancellations directly below for more details on billing.

Scheduling & Cancellations: The SRL online scheduling calendar is available for trained users to reserve instruments up to 30 days in advance. To make a reservation, users must have completed the required training module for that instrument, and be assigned to a valid funding source. For cell sorters, users are required to have an active biosafety form for the cell type they are sorting in order to make a sorter reservation; this form must be approved by the user’s PI before Penn Cytomics approval. Users may only schedule up to 3 hours of consecutive time on analyzers during normal operational hours (Monday-Friday, 9am-5pm). Longer sessions are acceptable during off-peak evening hours and weekends (see Access Policy). High-parameter analyzers (A5/SE Symphonies) and sorters are excluded from this policy, as it is recognized that these experiments generally require a significantly longer time to complete.

The following policies have been implemented in order to maximize instrument usage and to minimize placeholder reservations:

  • For most analyzers: The scheduling calendar prevents adding, modifying, or deleting instrument bookings within a 6-hour time window before a user’s scheduled time.
  • For A5/SE Symphonies: The scheduling calendar prevents adding, modifying, or deleting instrument bookings within a 36-hour time window before a user’s scheduled time.
  • For cell sorter reservations (trained users only): The scheduling calendar prevents adding, modifying, or deleting instrument bookings within a 24-hour window before a user’s scheduled time. Same-day cancellations for trained users/self-sorting, will be billed a minimum charge of 75% of scheduled time.
  • For staff-assisted sort reservations: All assisted sort reservations can only be booked/modified by calling the core. The SRL enforces a 72-hour cancellation policy; users are billed the entirety of the scheduled time, unless the SRL can fill that time interval with another reservation.
  • For training reservations: The SRL enforces a 24-hour cancellation policy for cytometer training reservations. All no-shows and late cancellations will be charged a cancellation fee (See Penn Cytomics website for fee amount).

*If a user needs to schedule, modify, or delete a reservation within these time windows, they will have to call 215-898-3528 to speak to a staff member*

Users may not “share” scheduled time with fellow lab members, as this will result in a double charge within the SRL billing system (User 1 is billed as a no-show, User 2 is billed for actual instrument usage). In this instance, User 1 should cancel their reserved time, and User 2 should reserve the instrument under their own credentials.

Data and Nomenclature: User data are stored on the local computer for 4 weeks after acquisition for analyzers and 3 months after acquisition for sorters. When users export data from the acquisition software, and upon completion of the usage session, Entrée automatically uploads data to two independent locations. Please use the following naming convention: PennKey_YYYYMMDD_Descriptor (ex: jonesder_20221103_Data). Do NOT use special characters, except for underscore (_) and dash (-). This convention allows staff to identify the owner of the data, indicates when data are eligible for deletion from the computer, and assists in future data retrieval requests.

Computers: The computers are for data acquisition and analysis only. Internet browsing is not allowed, as the computers are on a local network. In addition, USB flash drives are forbidden in any Penn Cytomics workstation computer, as to prevent the transmission of viruses and other malware.

Consumables Provided: Penn Cytomics supplies sheath fluid, 2% contrad, FACSClean, bleach, and BD detergent. Labware/reagents, such as tubes, filters, transfer pipettes, pipette tips, buffers, etc. are to be supplied by the user. Lab coats, gloves, and other PPE are the responsibility of the user. In satellite locations (areas other than JMB), the department is responsible for disposing of its own sharps containers, biohazardous waste, and trash. In satellite locations, chairs/furniture are also the responsibility of the department to replace.

SRL Policy Violations: The SRL employs a 3-tiered process to address failures to adhere to written policies. After a first-time offense, SRL staff will reach out to the responsible party to specify which policy was broken. If this user becomes a repeat offender despite staff intervention, their Principal Investigator will be notified. If the user still fails to adhere to policy, they risk suspension of privileges. Staff members will use their judgment on a case by case basis and may escalate this 3-tiered process based on the severity of the offense. For example, if a trained user logs in/allows an untrained lab member to operate any of our instrumentation, this will automatically result in their PI being notified and suspension of instrument privileges.

Acknowledgements: All publications that include results or products generated in the Penn Cytomics and Cell Sorting Shared Resource Laboratory must include the following acknowledgement:

“The data for this (manuscript or presentation) were generated in the Penn Cytomics and Cell Sorting Shared Resource Laboratory at the University of Pennsylvania and is partially supported by the Abramson Cancer Center NCI Grant (P30 016520). The research identifier number is RRID:SCR_022376.”