RII DIVISION FLOW CORE

User Resources & Guides

General Usage Guides

  • Turn on and log into Windows 10

  • Open Tera Term application on desktop and leave running in background

  • Log into Diva before turning on the instrument

  • Check liquid tank levels, refill if needed

  • Empty waste tank making sure to not get the filter wet. Refill with bleach to first marked line on outside of tank so that a full tank is > or = 10% solution

  • Check bubble filters on wet cart for air pockets. If air presents, user might have to get rid of all bubbles prior to proceeding

  • Give the lasers 30 minutes of warm-up time

  • You MUST filter samples immediately prior to loading on machine using 70uM mesh

  • Add EDTA to prevent clumping (when possible)

  • Use a concentration that does not exceed 15k events/second when acquiring at Medium

  • Run 10% Bleach solution every 20-30 samples for 5 mins, then dH2O to prevent clogging

Frequently Asked Questions

Troubleshooting

Instrument Discounted, Multiple Errors in Diva

Cycle power on the computer and cytometer

  1. Power down the cytometer
  2. Shutdown the computer (no restart)
  3. Let sit for 30 seconds
  4. Turn on the computer first and allow for a full boot up and log in to Windows with PW
  5. Open Tera Term and log into Diva
  6. Power on the cytometer

Air in flow cell or tubing

Fortessa

  1. Place tube of water on probe and place cytometer in STANDBY
  2. Make sure the wet cart power button is on
  3. Make sure the platinum tank has liquid in it and the wet cart has full tanks and no alarm light on (RESET button on wetcart if so)
  4. Hit the Prime button on the wet cart to refill the platinum tank while bleeding air filter and line above filter on line going in to side of machine
  5. Move the sample arm to the left and REMOVE water tube
  6. PRIME 2x with no tube loaded and arm to the side
  7. RUN water tube for 1 min on HIGH
  8. Try sample tube again
  9. Repeat above steps 2-3times, if this does not fix issue contact Core Manager

Canto II

  1. Check that no air is in the air bubble filter on the wet cart (if so, bleed air bubble filter)
  2. Click Cytometer Menu->Cleaning Modes->Prime Tank After Refill->Check all 3 boxes->OK
  3. Click Cytometer Menu->Cleaning Modes->Bubble Filter Purge & Degas Flow Cell
  4. Repeat Step 3, 2-3 times
  5. Run samples
  6. If not fixed, contact Core Manager

Clog

*users must filter all samples immediately before running on cytometer using 70um mesh filter or less

Fortessa

  1. Load a tube of 10% Bleach and mark meniscus
  2. Run for 2-5mins on HIGH
  3. If no liquid movement, unload tube PRIME2x, and reload tube repeating step 2
  4. If no movement or back pressure is too high, contact Core Manager
  5. If liquid level drops and clears the clog run water for 2-5 mins

 

Canto II

  1. Load tube of water and mark meniscus
  2. Click Cytometer Menu->Cleaning Modes->Clean Flow Cell
  3. Repeat step 2, 2-3 times
  4. If no liquid movement, load tube of Bleach and run for 5mins on high
  5. If still no movement, contact Core Manager
  6. If liquid level drops, clean flow cell 2-3x with water and then run water for an additional 2 mins and wipe down the probe

RII1 Data Storage Server Access

Email Howard Katz for ACI members or I-Cheng Ho for RII members to request access to RII1.

Once you have access: On the Windows desktop of a FACS computer, double-click the icon labeled BWH-RII1 (if it’s missing, see instructions* below)

A login box should pop up.  Enter your Partners user name in this format: partners\username, and enter your Partners password.  (NOTE: If a folder opens rather than a password box, and it’s not your lab’s folder, see instructions** below to change to your lab’s folder, or just use the folder named “FACS Data” to transfer your data.  The FACS Data folder can be accessed by all users of RII1.)

Click OK

Your lab’s folder should pop up in a window.  Copy your files from the FACS computer to RIA3 as you would between any two folders in Windows.

To log off your folder, right-click on the “My Computer” icon on the Windows desktop.

Select “Disconnect Network Drive”

Select \\rfawin.partners.org

Click OK

Contact Howard or I-Cheng if it doesn’t work.

See separate instructions for how to access your RIA3 folder from other computers.

 

* If desktop BWH-RII1 icon is missing

Right-click the “My Computer” icon on the desktop or in the Start Menu

Click “Map network drive” to open the dialog box

In the “Drive” field, select drive “Z” from the drop-down menu

In the “Folder” field, enter this address: \\rfawin.partners.org\BWH-RII1

Be sure that “Reconnect at logon” is checked

Click on “Connect using a different user name”

Enter your Partners user name in this format: partners\username (note that it’s a back slash, not a forward slash), and enter your Partners password

Click OK and Finish

Double-click the “My Computer” icon, and you should be able to right-click on the letter of the drive you just mapped (i.e., “Z”)

Select “Create shortcut”

Click yes to the request to put it on the desktop

The RII1-bwh icon will appear on the desktop.  You can shorten the long name Windows will put under it by right-clicking on the name and shortening it to RII1-bwh.

**If RII1 is logged into someone else’s folder

Click on the “My Computer” icon on the Windows desktop, or in the Start Menu

Select “Log Out”

Re-log into Windows with username BDOperator

Data Management

  • Experiments containing data are not allowed to be stored within the Diva software or on the cytometer computer. They must be stored on the RII1 server.
    1. Export all experiments containing data to RII1 servers immediately after run
    2. If you’d like to store a blank template in the software for future use: Right-click the experiment and “Duplicate Without Data”
    3. Delete experiment containing data and save template for future experiments
  • When exporting, export data as both an “Experiment” to backup Diva software settings and as “FCS Files” for analysis using 3rd party software on your personal computer

Booking Rules and Regulations

  • Each user must complete our full Orientation & Training process to gain access to the core
  • Each user is required to have an individual log-in and cannot use a general lab account
  • Users are allowed to book up to a maximum of 2 hours at a time during “peak hours” (Mon-Fri 12-5pm)
  • Each user is responsible for making sure the instrument is shutdown or passing that responsibility on to the next user on the calendar.
  • If you are the last user of the day, it’s your responsibility to shutdown the instrument for the day, or to contact the 2nd last user of the day or core personnel to shutdown the instrument. Users who are unable to follow rules will be banned from access for 3 weeks.

    CANCELLATION POLICY

    1. Users must cancel unused bookings 6 hours before the start time of the event to avoid charges
    2. If a user cancels their booking 6 hours within their start time, and the vacated time goes unused, the user will be charged
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