The IGC-FCF is open to IGC researchers and to investigators outside IGC.
New users must first register in our booking system, Agendo, and then request permission to gain access to the calendar of the desired equipment.
To use the FCF services, users aiming to do flow cytometry analysis must enroll in our Training Program, consisting of one theoretical and one hands-on module. This training is optional for those users only doing cell sorting.
Users may be charged for assistance if needed.
All users must read, understand and agree with the policies described here. Any questions should be directed to the IGC-FCF staff.
It is mandatory to reserve through Agendo any equipment before usage, including weekends.
To gain access to instrument calendars, users must request permission for the desired equipment through Agendo.
Using equipment without reservations is forbidden and will be reported to the Head of FCF and to the IGC Direction as misconduct.
An instrument can be booked up to 14 days in advance in blocks of 30 minutes (analyzers) or one hour (sorters).
Booking of consecutive slots of time for the same user is not allowed. Users must make a single reservation for the entire time period intended.
Experiments are scheduled on a first-booked-first-served basis.
Filling all reservation fields is mandatory, and must be in accordance with the field's title. All requested information is necessary for assistance and/or statistical purposes, to help improve the quality of the service. Failure to fill adequately the requested fields will be considered improper usage and may lead to reserving limitations for the abusing user.
Reservations for analyzers are automatically confirmed upon user login in Agendo on the workstation connected to the respective equipment, within a specified period of time before, during, or after the reservation time. Failure to do so indicates the user did not use his/her reservation slot, being automatically reported by e-mail to the user and to the FCF staff.
All the reservations are charged, unless they are deleted.
If the user decides not to use the equipment, the reservation must be deleted at least four hours (analyzers) or 48 hours (sorters) in advance.
When deleting a last reservation of the day, investigators must ensure that the previous user and the FCF staff are both notified.
Users must be aware that even if their reservation is not the last one of the day, it may become so if subsequent slots of other users are deleted meanwhile. Therefore, every user must check the Agendo calendar before leaving the analyzer’s room, to ensure he/she has not become the last user of the day.
The full responsibility for the proper usage of the equipment belongs to the user who has the reservation in his/her name.
Reservations for the cell sorters work as pre-reservations and require confirmation from the FCF facility according to staff availability.
Filling all reservation fields for the sorting pre-reservation is mandatory to help FCF staff do determine the best instrument, configuration, settings and sorting time.
Approval of and/or changes to the pre-reservation will be notified by e-mail.
It is the responsibility of the PIs to ensure their students and postdocs comply with all the policies described here.
Cell Sorters – FACSAria IIu, MoFlo
The instruments and operators are available for sorts Monday through Friday, from 10h00 to 18h00, except during maintenance periods (see Section Weekly Maintenance below).
The sorts are scheduled on a first-booked-first-served basis.
Cell sorting scheduling is done at the discretion of the FCF, to maximize availability for the entire IGC community. FCF staff schedules sorts preferentially from the beginning or at the end of the day to avoid breaking large time blocks.
If time is unscheduled one day prior to sort, FCF staff can adjust the schedule to fit better particular user needs.
To schedule a sort, please:
Check time availability for the intended instrument in Agendo
Fill up all the fields required upon Agendo booking
Get follow-up confirmation e-mail from FCF staff
Prior to the first sort of a new protocol, please arrange an appointment with FCF Head to discuss the details of the sort.
All the sorts are performed by FCF operators, unless users choose to attend the FCF training module of Cell Sorting and acquire sufficient experience to become ‘Power Users’ (option available for FACSAria only).
It is mandatory to decontaminate the flow cytometer analyzers immediately after sample acquisition, following the cleaning procedures indicated by the FCF. Detailed notes on washing procedures are posted on the instruments.
After decontamination, the cytometer must be switched to STAND BY and stay with a tube with H2O installed.
It is mandatory to refill the PBS container at the end of acquisition.
Instruments, computers and screens should be turned off if nobody is scheduled to use it in the following four hours and after the last user of the day.
The last user of the day is fully responsible for shutting down the equipment. Failure to do so may incur in penalties for the user and costs for the PI for any damage directly related to this serious misusage.
It is strictly forbidden to remove the Sample Injection Port (SIP) outer sleeve from any instrument. If you believe you need to do so, please contact the FCF staff.
If the user experiences any unusual behavior with the cytometer and/or during the analysis, he/she should first refer to the Troubleshooting section of the User’s Guide that is available next to each instrument.
If the situation cannot be solved according to the Troubleshooting recommendations, the user should call the facility's staff for assistance.
It is the user’s responsibility cleaning up the working bench space and discard all the tubes into the green containers for biohazard waste after his/her experiment.
Analysis Workstations – Folk Workstation (PC), Ohpah Workstation (Mac)
The computers are available 24/7.
Access to the computers is on first-come-first-served basis (without signup).
These computers are dedicated to flow cytometry data analysis only. They are on the IGC network, run Flow Cytometry software (FlowJo, BD FACSDiVa, FCS Express, ModFit, Summit, Flowlogic, Fiji, Milliplex Analyst) and Office programs to analyze and publish data.
The local hard disks of the analysis workstations are not for long-term data storage.
Users are responsible to make archival copies of their data.
The local hard disks of the cytometer workstations are not for long-term data storage.
All files are routinely (monthly) erased to maintain enough free disk space for acquisition and/or analysis, including data on FACS Diva.
It is strongly recommended to export experiments from the FACS Diva to the local folder immediately after acquisition, and transfer data from there to the user’s personal directory in the remote server.
The local folder that should be used to store temporarily data exported from FACS Diva is located in the Desktop (Export – Routinely Cleared). Any files or folders outside this folder will be deleted without notice.
The use of USB pens or external drives is not allowed.
The IGC-FCF is not liable for any loss of data on the local hard disks.
Regular weekly maintenance on all FCF Sorters, Analyzers and MAGPIX is scheduled on Monday or Tuesday mornings (see time scheduled in Agendo).
Regular weekly maintenance includes the following procedures on all FCF Analyzers:
System flush according to BD-recommended procedures for Weekly or Monthly Maintenance (BD instruments); extended acquisition of cleaning solutions & preparation for aseptic sorting (FACSAria); extended acquisition of cleaning solutions (MoFlo).
Autoclaving of FACSAria sheath tank (once a month after 16h00).
Quality Control according to BD-recommended procedures (all instruments).
Reposition of all cleaning solutions, PBS and dH2O, and 5-mL FACS tubes.
Removal and discard of all unlabeled sample tubes and racks from the desks and benches.
Purge of all illegitimate files from the computers (at the beginning of every month).
To recover essential operating costs, the FCF charges hourly fees for the sorts and use of the flow cytometer analyzers and MAGPIX.
A fee for each FlowJo license is charged annually to groups requesting these licenses for their personal computers.
Usage of cell counters is free of charge, but scepter sensors needed for cell counting are charged.
Usage of analysis workstations (other than those attached to the instruments) is free of charge.
Scheduled operator-assisted services (sorting and MAGPIX) must be canceled no later than 48 hours prior to the scheduled experiment. If the scheduled sort is not cancelled by this time, the booked time will be fully charged, even if the investigator decides not to use it.
Scheduled use of Analyzers must be cancelled no later than four hours in advance of the scheduled start time. If the scheduled use of the analyzer is not cancelled by this time, the booked time will be fully charged, even if the investigator decides not to use it.
If investigators do not cancel their booking within the free-of-charge tolerance period, but find someone willing to use the time they had booked, the FCF staff will transfer that booking to the name of the second investigator, so it is charged to the new user.
The subjects reviewed at the training sessions may vary depending on the background, needs and interests of the investigators. See FCF Training Program. Please do not hesitate to contact FCF Head to find out the best solution for you.
The Theoretical Training sessions are scheduled on a regular basis (check the calendar here); the Practical Training sessions are scheduled with the FCF staff on a first-booked-first-served basis after attending the Theoretical Training module.
For consultation and help, please contact directly the FCF staff during regular working hours or 24/7 via e-mail to get immediate feedback or schedule a meeting with the head of the facility.
By using the IGC Flow Cytometry Facility services, investigators commit to refer the Facility in all publications that include data generated by its services and/or resources in the Materials and Methods and/or in the Acknowledgements section. You are welcome to use the following examples of references:
In Materials and Methods: “All Flow Cytometry experiments were performed at the Flow Cytometry Facility of Instituto Gulbenkian de Ciência, Oeiras, Portugal”.
In Acknowledgements: “We would like to thank the Flow Cytometry Facility of Instituto Gulbenkian de Ciência for their services and assistance”.