Access to equipment Heading link
The Flow Cytometry Core Lab (FCC) is a pay per use service that provides access to state-of-the-art instrumentation. In order to access the instruments all users must be registered with the Research Resources Center, and have an active RRC Account Number (RRCAN) tied to a funding source which is usually a C-FOAPAL number but can also be a credit card or purchase order. This is mandatory to have access to the FCC. For bench-top analyzers, users have an option of having their samples run by trained research specialists (staff-assisted) or to train to run the samples themselves (self-run).
Only users who have undergone training by the FCC staff will have access for running the instruments themselves.
Only users trained on a specific instrument can access that instrument.
The instruments are available 24×7 for trained users who have created a history of good and reliable practice. Newly trained users will be granted access to the instruments only during regular business hours, between 9am and 5pm during weekdays. Once the user has created a positive history, FCC staff may determine to provide user 24×7 access.
It is mandatory that all users must wear a dedicated lab coat, gloves and protective shoes for all experiments performed in the FCC or using FCC instruments. Other protective clothing/material such as protective goggles may also be worn based on the users’ discretion and kind of experiment.
Training Heading link
Users wishing to train to run samples themselves will be provided a one-on-one training by the Director of the facility or one of the FCC staff.
- Training will be provided by FCC staff ONLY. Training received at previous institutions or provided by colleagues will not qualify the user to run the instruments at the FCC.
- The choice of instrument for training or assisted runs will be made based on instrumentation requirements based on experimental design. All the flow cytometers have multi-parametric capability with at least 3 lasers and the ability to read at least 9 different fluorophores. FCC staff will work closely with the users to determine the appropriate instrument for training. In the event of a future possibility of expanding the number of fluorophores or adding a fluorophore than can only be read on a specific instrument, users will need to undergo training for that specific instrument.
- All flow cytometers vary in their hardware and software configuration. Users wishing to train on different instruments will have to train for each of them separately.
- Training usually comprises two hands-on sessions of 90 minutes each. This includes a first introductory session to some basics of flow cytometry, an introduction to the acquisition software and actual run time on the instrument. The second training session includes supervised time where the user gets an opportunity to create protocols and run their own samples independently, rules of compensation, and an introduction to the importance of gating and some points on basic analysis of data. More detailed training for analysis will be provided in a separate module. Trainees are required to bring 5-6 samples including controls for the training. FCC staff will determine if more training is required beyond the two sessions for the user. The final decision to determine the readiness of the user to run samples independently and thereby gain access to run the instrument will be made by the FCC Director.
- The cell sorter is run by specialized staff only and training is not provided for the cell sorter.
- Users found accessing instruments for self-run without training will be barred from any future use of the instrument and all samples in the future will be run as staff-assisted only. In addition, a penalty of $300.00 will be levied against the user and charged to the PIs RRCAN. Under no circumstances shall one user train another user, regardless of their experience to run the instruments. The user providing such training will be penalized $300.00 and the respective PIs account will be charged. The user may also lose future rights to run the instruments.
While we strongly encourage investigators to use the FCC and are committed to providing cutting-edge technology and expert support, the use of FCC facilities is both a privilege and a responsibility. We are confident that users will go through proper training prior to using the instruments and will practice proper lab and ethical practices after having gained access to the instruments.
Reservation/Cancellation Heading link
All bench-top flow cytometers and charged in 5-minute increments with a minimum run time of 15 minutes. Cell sorters are charged in 30-minute increments. The reservation/cancellation policies vary with the instruments. Please ensure that you have read the reservation/cancellation policies for each instrument to avoid unnecessary penalties. All users who require staff-assisted runs will be provided access to the instruments on the calendar without the need to train. For self-run users, training is mandatory before receiving access to make reservations on an instrument.
Reservations Heading link
- Users are charged for the entire duration of reserved time.
- Users are expected to be punctual and use the instrument during their reserved time only.
- In the event, that the user exceeds the reserved time, a new reservation should be made for the additional time.
- Users who have made a reservation will have first right of access to the instrument for the particular time slot.
- Users who have not made sufficient time and have unfinished samples at the end of run must give right of access to the next user with a reservation.
- Users MUST ‘Begin reservation’ on UICore prior to using the instrument and ‘End reservation’ after disinfecting and cleaning the instrument. Reservations that have no start or end time require FCC staff to investigate the logs to determine start and end time. Users will be charged an additional 15 mins of staff-assisted time on the reservation time for this effort. In case of any internet glitches with the internet or UICore, please inform FCC immediately with the actual run times to avoid unnecessary penalties.
- For the Cyan 1 located as a satellite facility on the 8th floor, beginning reservation on UICore will turn on the monitors thereby giving Users access to the instrument. Any effort to override the relay mechanism will result in the user losing privileges to run the instrument with immediate effect.
- A single reservation cannot exceed a 4-hour period. In case the time of run required exceeds 4 hours, permission from the Director must be acquired to make successive reservations.
- Repairs and breakdowns are an unfortunate reality for any instrumentation. In case an instrument breaks down, FCC staff will either try to fix the instrument or provide an alternative instrument to accommodate samples. If these options do not work, then users will be requested to fix samples to be run at a later time after the instrument has been fixed. Under such exceptional circumstances, it may be necessary to move all reservations down until the issue is resolved. Reservations will be moved down in the order they were made.
- Users will be given credit for time lost due to a broken instrument. This does not include time lost due to clogs resulting from poor sample preparation or misuse of the instrument or an error by the user resulting in the breakdown.
- In case of instruments that are not working, please inform FCC staff immediately. Do not attempt to fix or repair the instrument yourself. This may result in damaging the instrument further.
Cancellations Heading link
Cancellation policies vary with the instrument.
Cancellation of self-run or staff-assisted runs before 48 hours of the run on the Cyan and Gallios will not incur a penalty. However, a cancellation made between 24 and 48 hours will result in a $10 cancellation charge for the Cyan and Gallios. Cancellations made the same day will result in full charges for the time of reservation.
Cancellation of self-run or staff-assisted reservations on the LSR Fortessa must be made at least 72 hours before the run to incur no penalty. Cancellations made between 48 and 72 hours will result in a $10 cancellation charge. Cancellations made within 48 hours will incur full charges for the time of reservation.
Cancellation of cell sorting reservations on the MoFlo Astrios made at least 72 hours before the run will incur no penalty. Cancellations made between 48 and 72 hours will result in a $100 cancellation charge. Cancellations made with 48 hours will incur full charges for the time of reservation.
Cancellation charges may be waived if the time is used by another user.
A ‘no-show’ without prior intimation may result in cancellation of the reservation 15 minutes after the start time. FCC will decide if the time can be given to another user to run samples. In case there is a delay in bringing samples, please call the flow cytometry lab to inform them to avoid losing your reservation.
Cell sorting Heading link
- For cell sorting, uses are required to call the FCC lab to make the reservations. Information about the fluorophores that will be used for the experiment should be provided at the time of making the reservation. Sorting requires the lasers to be aligned and quality control run for each laser, prior to sorting. Only lasers required for the specific fluorophores are turned on, aligned and assessed for QC. Users are request to avoid last minute surprises as it may cost time to align the lasers and may result in insufficient time to finish the sorts.
- Please make sufficient time to ensure to complete your sorts. Users that have made reservations will have first access.
- All samples should be filtered using a cell strainer prior to sort. Users will be required to provide the tubes. In case the users do not do so, FCC will provide for the tubes and charge $2 per tube.
- A good range for sample concentrations is generally between 10-20 million cells/ml. Users should bring some sample buffer in order to dilute the cells if too concentrated.
- If there are any specific instructions regarding speed of run, nozzle size requirements, gating strategies and requirement of purity testing, please explain this to the research specialist well before.
- Clonal sorts require additional time for set up. Please inform the research specialist running the sorter well in advance about your requirements and make adequate time for the same. Give at least 30 minutes for clonal sort set-up time.
- In the eventuality of problems with the cell sorter, all reservations will be moved down according to the order in which they were made. This does not include time lost due to clogs resulting from poor sample preparation or poor sample signal or lack of proper controls.
- Biohazardous samples will not be run without adequate fixation of the samples to ensure that the samples are completely disinfected and safe for run. The FCC staff will reserve the right to refuse running or sorting samples that can be potentially biohazardous.
- It is assumed that all investigators running animal, human or cell line samples have adequate approved protocols for the experiment. ACC/IRB protocol numbers must be filled in the sample sheet at the time of sort. FCC can refuse to run samples if this information is not provided. It will be Principal Investigator’s responsibility to ensure that these protocols are in place. FCC will not bear responsibility for any violation of University protocols/ethics codes on the part of the users.
Data acquisition Heading link
- All self-run users should fill the sheath buffer, empty waste buffer prior to running the instrument. Instruments should be thoroughly disinfected by running 10% bleach and then cleaned with detergent and water as instructed during training.
- All samples should be filtered with a cell strainer prior to running on bench-top analyzers. Users will be required to filter the samples. In case the users do not do so, FCC will provide for the tubes and charge $1.75 per tube.
- No bio-hazardous samples should be run on the bench-top analyzers. (Please see University policies on the definition of bio-hazardous material).
- All human and animal tissue and cell lines must be fixed prior to run.
- It is assumed that all investigators running animal, human or cell line samples have adequate University approved protocols for the experiment. ACC/IRB protocol numbers must be filled in the sample sheet at the time of acquisition. FCC can refuse to run samples if this information is not provided. It will be Principal Investigator’s responsibility to ensure that these protocols are in place. FCC will bear no responsibility for any violation of University protocols/ethics codes on the part of the PI/User.
Data transfer and archiving Heading link
- Data acquired on the bench-top analyzers and cell sorters are transferred to the My Data server via a software script. This process usually takes a few hours. In case the data is required immediately, please bring a CD or DVD to copy the data from the computer. Use of Flash drives including ‘brand new ones’ is strictly prohibited. This is to protect the computers from viruses which may otherwise result in corrupting the software and render the instrument unusable.
- All self-run users are expected to transfer the data themselves. Neither the software nor the computers connected to the instruments are meant for archiving data. Large data can slow down the software and can even crash the computer resulting in long down times.
- Once data has been transferred to the My Data server, the data will be removed from the software immediately and from the computer within one month of acquisition without notice. This should give sufficient time for the users to confirm that they have received and saved the data.
- Please ensure to name your FCC folder in the format e-mailID-4numbers(
usually month and date)-textwithoutspacing. Folder has to be zipped and loaded to the My Data drop folder on the desktop, incorrect format can result into no data. The files need to be downloaded from your account ASAP. All the data past 60 days will get deleted.
- FCC will not be responsible for data not received or lost beyond these periods.
Use of FCC computers, Do's and Dont's Heading link
- No new authorized or unauthorized software should be downloaded on the computer.
- FCC computers should not be used to watch videos, shop or surf the internet.
- Use of flash drives is strictly prohibited.
- You are welcome to bring a CD or DVD to copy the data from the computer.
Acknowledgement of Use of FCC Heading link
We strongly encourage users to acknowledge the use of FCC facilities in your publications. You may use the following text for general acknowledgment-
“This work made use of facilities in the Flow cytometry Core lab (Research Resources Center, UIC)” If there are specific staff you would like to acknowledge please do so.
Do's and Don’ts Heading link
- Do wear protective lab wear for your biosafety.
- Do begin and end your reservation in a timely fashion.
- Do fill the sheath tank prior to run and empty the waste tank after the run.
- Do disinfect and clean the instrument after your run.
- Do filter your samples before running them.
- Do send your data via the My Data server after run.
- Do talk to FCC if you have any questions regarding design of the experiment.
- Do acknowledge FCC in your publications.
- Don’t be late for your appointments.
- Don’t run biohazardous samples without appropriate fixation.
- Don’t run samples without suitable controls.
- Don’t use the computers to watch movies, shop or browse.
- Don’t assume that all flow cytometers will detect all fluorophores.
- Don’t lose your patience if there are issues with the instrument.
- Don’t assume that your data on the instrument will stay there forever.
- Don’t use flash drives on the computers.