IGC

Equipment

The Flow Cytometry Core Lab currently has 5 flow cytometers divided in two categories.
  1. Bench Top Analyzers: These are used only for acquisition, and are operated by researchers themselves. Reservations are available here.
  2. High Speed Cell Sorters: Their prime use is for cell separation, and dedicated operators within the facility staff are required. Exceptions are possible both for acquisition and for self-operation of the instruments, based on experience and approval by the facility's staff. Reservation is done by sending an email to the Facility's staff.

Our instruments are equipped with several laser lines summarized in the following table. The instrument to use is chosen based on the panel of fluorochromes to be detected, the optical configuration to detect those fluorochromes, and on the availability of the machine. To find out more about each instrument's optical configuration, see individual descriptions below.

Bench Top Analyzers

There are two Becton Dickinson (BD) bench top analyzers available, that can detect up to 3-4 colors simultaneously. Daily cleaning, in addition to weekly complete fluidics cleaning and quality control of the optical system are performed in both analyzers (Reports available soon). Each investigator should be aware of the startup and shutdown procedures, as well as the cleaning procedures after usage. If there are any doubts please ask for assistance, and make sure you have read the facility's policies on instrument usage.

  • FACScan
  • BD FACScan | Reservations

    There are two Becton Dickinson (BD) bench top analyzers available, that can detect up to 3-4 colors simultaneously. Daily cleaning, in addition to weekly complete fluidics cleaning and quality control of the optical system are performed in both analyzers (Reports available soon). Each investigator should be aware of the startup and shutdown procedures, as well as the cleaning procedures after usage. If there are any doubts please ask for assistance, and make sure you have read the facility's policies on instrument usage.

  • BD FACScalibur | Reservations

    This is a 2-laser 4-Color flow cytometry Analyzer produced by Becton Dickinson.

    Up to 3 colors can be detected from the aiircooled 15mW blue (488nm) argon laser (e.g., FITC, PE, and PerCP) and another color from the red (635nm) diode laser (e.g., APC or Cy5). It's also equipped with a direct waste system eliminating the need for a waste tank. The instrument is connected to a Power Machintosh G4 loaded with BD CellQuest for machine interface and data acquisition.

    More information on BD FACSCalibur...

  • FACSCalibur
  • CyAnADP
  • BC CyAn ADP | Reservations

    The CyAn ADP is a 3-laser 9-Color flow cytometry Analyzer (BC) manufactered by Beckman Coulter.

    It's equipped with a 25mW solid state blue (488nm) laser, a 50mW solid state violet (405nm) laser, and a 60mW red (642nm) diode. It is able to detect up to 9 colors simultaneously: 5 from the blue laser (e.g., FITC, PE, PerCP, PE-TexasRed, PE-Cy7), 2 from the violet laser (e.g., Pacific Blue and Pacific Orange), and 2 from the red diode (e.g., APC and APC-Cy7). The instrument is connected to a PC loaded with Summit Software for machine interface and data acquisition.

    The CyAn ADP is equipped with the HyperCyt autosampler, which allows automated sampling of multiwell plates.

    More information on BC CyAn ADP...

  • Fortessa
  • LSR Fortessa SORP | Reservations

    LSR Fortessa SORP is a powerful bench cell analyzer from Becton-Dickinson, equipped with 3 lasers (violet, blue and yellow flowcytometry), detecting up to 13 colors simultaneously.

    Flow cytometry data is analyzed using BD FACSDiva™ software for an efficient setup, acquisition and analysis. Analysis can be made in tubes or in 96 well plates, using BD™ High Throughput Sampler (HTS).

    More information on LSR Fortessa SORP...

  • Cell Sorters

    There are two High-Speed Cell Sorters available, that can detect up to 8-9 colors and sort up to 4 populations simultaneously. Both instruments are able to perform multiple or single cell sort onto standard 96-well plates or any custom plate. Sorting speed in both machines can theoretically go up to 100,000 cells/sec, though depending on sample type, population frequency, and normal sorting constraints, sorting rates are usually <10,000 cells/sec. Routine fluidics cleaning and quality control of the optical system are performed regularly in both sorters (Reports available soon). Both sorters can be used for acquisition when detection of at least one of the fluorchromes is not possible in the available analyzers. Due to the level of complexity in handling the MoFlo, analysis is always assisted by the facility's staff in this sorter. FACSAria can be operated for analysis by users that have undergone some type of training, and have been approved by the facility's staff.

  • BC MoFlo | Reservations

    MoFlo is a high-speed cell sorter produced by Dako Cytomation (now owned by Beckman Coulter), highly configurable and adaptable to the researcher's needs.

    MoFlo stands for Modular Flow Cytometer, since it's built on individual modules, which can be configured to basically anything from a simple 1-laser 4-color Analyzer to a 3-laser 8-color High-Speed Cell Sorter. Our Model is configured with up to 9 PMTs (8 Colors), 4-Way Sorting, and the Cyclone Single Cell Cloning Unit. We have custom modified it to accomodate a refrigerated 4 Way Collection Station, custom made dichroics holders, an alternative fluidic system using 2 separate PBS and dH2O cleaning systems, different fluidic assemblies for sorting microorganisms or higher eukaryotic cells, and a custom drop-delay measurement system based on BD's accudrop system.

    Excitation sources include a 200mW Sapphire 488nm (blue) laser, a Coherent I 90-4C 4Watt Ar-Ion Laser (MLUV and several visible lines), a Spectraphysics 35 mW HeNe 635nm (red) Laser, and a custom mounted 50mW DPSS 561nm (yellow) laser connected through fiber optics. However, only 3 of the 4 lasers can be used simultaneously. MoFlo is a jet-in-air system, and consequently sensitivity or excitation efficiency is lower than with cuvette-based flow cells (such as with FACSAria). However, this can be overcome by using higher power lasers, though some fluorochromes, such as PerCP and its conjugates, are still not detected efficiently in this instrument. Only dedicated staff are allowed to operate the machine.

    More information on BC MoFlo...

  • MoFlo
  • FACSAria
  • BD FACSAria | Reservations

    FACSAria is a high-speed sorter manufactured by Becton Dickinson. It's equipped with three low-power air-cooled lasers at 488-nm (blue), 633-nm (Red), and 407-nm (violet) wavelengths enabling multicolor analysis of up to 11 parameters (9 fluorescent markers and 2 scatter parameters), simultaneously.

    With a fixed-alignment cuvette flow cell, fixed-optics design, and digital electronics, FACSAria is a user-friendly instrument, that can be operated with minimal training. The instrument is connected to a PC workstation loaded with BD FACSDiva software for machine interface and data acquisition.

    More information on BD FACSAria...