IGC

LSR Fortessa SORP | Reservations

Optical configuration

  • The LSR Fortessa is equipped with 3 different low-power lasers that can be used simultaneously. A 50mW solid-state blue (488nm) laser, from which SSC and another 5 colors can be detected (upgradable to another 2); a 40mW yellow-flowcytometry (561nm) laser, from which 5 colors can be detected; and a solid-state 40mW violet (446nm) laser, from which 2 colors can be detected (also upgradable with an extra color). Instead of calling the detection channels generically FL1-FL9, BD named each channel after the most typical fluorochrome it detects. Note that a band-pass filter designated as 530/30 indicates that it will detect light in the a 30nm range centered at 530nm, i.e., 515-545nm. LP stands for Long Pass filter, and indicates that it will detect light above the mentioned wavelength.

Operating Procedures

  • Before Acquisition

    1. If machine is turned OFF, follow the Startup LSR Fortessa procedure.
    2. Check if sheath tank is full. If necessary refill tank with PBS.
      Clean up all spills. PBS is corrosive!
    3. Start BD FACSDiva and log in as Administrator (no password needed).
    4. Wait for the cytometer to connect to the computer.
    5. Remove tube from Sample Injection Port (SIP) and PRIME 3 times (with support arm to the right).
      Check Bubble Filter and flow cell for bubbles. If necessary change the connection tube to the orange quick-connect and slightly tap the filter while opening the bleed valve.
    6. Run PBS for 1 min in HIGH to check if machine is dirty. If necessary follow the Cleaning Procedures.
      Though events may appear, it may not be due to poor cleaning and they may or not interfere with acquisition. Evaluate noise events based on the location and event rate during sample acquisition.
    7. Select the right Configuration: Cytometer View Configuration, select the correct configuration and click Set Configuration.
      Make sure filters and mirrors on cytometer match the configuration you selected (if not the case, make the necessary changes – no slot should be empty!)
    8. Open/Create your Experiment and setup acquisition plots in Diva.
    9. If you are using the HTS set the button on the right side of the Fortessa to plate position, and in Diva software go to HTS → select Prime, 2 times.
    10. You are ready to start running your samples.
  • Startup Fortessa


  • After Acquisition

    1. Run Cleaning Procedures
    2. When finished, check if there is a user reserved after you in the next 3 hours.
      IF YES: leave the Fortessa on Standby with dH2O on the SIP and and refill the sheath tank.
      IF NO: Close Diva software (don't use the red cross, use File → Quit)
      Switch OFF the power extension connected to Fortessa and refill the sheath tank
    3. Export your acquisition files from Diva and REMOVE all your data from the software and hard drive.
      The computer is NOT for data storage.
      The hard-disk is subject to routine cleaning, and files may be deleted without warning.
      Suggestion: You may remove your files while running Cleaning Procedures (see instructions on how to transfer files to the xservers).
    4. Remove your gloves, tubes and other belongings and clean up any trash.
    5. If you have any doubts or need assistance call the facility's staff (ext. 251).
  • Cleaning Procedures Fortessa


  • Trasfering Data

    Acquired data is saved directly on the hard drive. Use the D:\facsdata\ partition for this purpose.

    The way data is exported, depends on where the data is to be analyzed:

    export as Experiment if you are going to analyze on an offline Diva software & wish to keep plots & gates

    export as FCS files to analyze data in any flow cytometry software

    To avoid reaching the limit size of Diva Database file, all Experiments must be exported after acquisition. Please confirm if files were successfully exported to your folder and DELETE them, leaving only an empty Experiment template within the software.

    Due to space constraints, all data is routinely cleaned. To avoid data loss, the users MUST transfer their data to another location. The best way to store your own data is to transfer it to a pen-drive, USB external hard disk, or to your directory in remote server. To transfer data to filespc or to another network location, follow these steps:

    1. Open any folder window.
    2. Write "\\filespc" on the location bar and hit Enter.
    3. You'll be prompted for your username and password.
    4. Select the folder you want to access.
    5. Locate and drag your local files into the remote folder.
    6. To disconnect from the remote drive you have to run one of the 'windows-logout' executable files located in the remote server.

  • Custom Changes

    Our LSR Fortessa has been custom changed to allow the inspection of the instrument’s flow cell, easily allowing the examination for the existence of bubbles in the flow cell and also laser alignment.

    custom_change

    The machine also includes a direct waste system eliminating the need for a waste tank. Therefore, users do not have to keep changing the waste tank when it's full. The sheath tank, however, must be ALWAYS refilled after usage, or if running low on PBS.