IGC

Cell Sorters

We have currently 2 cell sorters available, each with different assets. The cell sorters are operated by the FCF staff, and as such it is dependent not only of the machine availability, but also the staff availability. To schedule an experiment talk to the FCF staff.

List of Sorters and their configurations

BD FACS Aria IIu

BD FACSAria IIu is equipped with 3 different lasers that can be used simultaneously. It can sort up to 20 000 events per second with high precision.

It can sort 4 populations at the same time to eppendorfs, 5mL tubes or PCR tubes, and 2 populations at the same time for 15mL falcons. It can also sort to PCR strips, 6/96/384-wells plates, to terazaki plates or to slides (one population at a time). It is installed in a security chamber which allows us to sort human samples with biological risk (BSL2). It comes with 3 nozzles with different sizes (70, 85 and 100 µm) that we can change depending on the samples.


NOTE: Besides the fluorochromes mentioned above, we have filters that enable us to read BV605, BV711 ou BV785. Be aware that Aria can only read 2 colors in Violet simultaneously. If you need to use these colors, contact the FCF staff.

MoFlo

Moflo is equipped with 4 different lasers, although the Multiline (514 nm) and the Yellow Green (561 nm) lasers can't be used simultaneously.

It can sort 3 populations at the same time to epperdorfs and 5mL tubes. It can also sort to 6/96-wells plates or to slides (one population at a time). It comes with 2 nozzles with different sizes (70 and 100 µm) that we can change depending on the samples.


Pricing

Type
Price/hour
Internal
43.99 €
Academic
43.99 €
Campus
43.99 €
Industry
88 €

The use of the sorters is paid per hour of reservation.


General Policies For Sorters Usage

  1. It is mandatory to have a reservation in Agendo to use the cell sorters.
  2. Investigators must fill all the reservation fields in Agendo correctly because FCF staff need to know which lasers to set up and what optical filters to use.
  3. If you would like to book a sorting for the first time or in the context of a new experimental approach, please arrange a meeting with the head of the FCFto devise a plan and receive advice on how to prepare your samples for sorting, which instrument to choose, time length of your booking, etc.
  4. Every reservation must be confirmed by the FCF staff to ensure there is an available person to operate the cell sorter.
  5. It is mandatory to filter all samples and controls to be analyzed in the cell sorters to ensure clumps are removed. This filtration step must be performed in the Flow Cytometry Lab just prior to sample load into the instruments. The IGC-FCF can provide pipettes, tips and filters when needed, but you are welcome to use your own filters.
  6. In the day of cell sorting, the FCF recommends investigators to bring:
    1. Unstained control
    2. Single staining controls
    3. Viability staining
  7. The ideal cell concentration for cell sorting is 5 x 106 cells/mL (for large and/or sticky cells) or 2 x 107 cells/mL (for small cells). For single-cell sorting, sample concentration should not exceed 1 x 106 cells/mL.
  8. Bring the collection tubes/plates coated with protein.
  9. Bring an organized gating strategy already well thought.
  10. Please consult the Preparation for Cell Sorting Guide here. You can also get a pdf with instructions here.

More information regarding facility policies here. Any missing information, contact the FCF staff.