Getting Started

Information About Our Facility

Availability

In order to sign up to use machines, you have to be formally trained by the facility. Facility notifies users of the webpage, in which you agree to all its policies pertaining to the proper use of the instruments. You cannot train others in your lab to use the instrumentation. Once you are trained and allowed access to the calendar, you can book time online.

  • The CytoFLEX LX, Aurora, Gallios, and the Celigo are available 24 hours/day, 7 days a week.
  • All users are required to sign in on the on-line calendars before using the instrumentation.
  • To access these machines, post training access, card programming, and authorization forms are available from the flow facility.

Security

You need a Proxy card to gain access to the Flow Core Facility. After receiving authorization from the Flow Core, you can get a Proxy card through the Public Safety Office or have your current Proxy card activated.

For questions or concerns, please contact John Tigges at 617-735-4191 or email at jtigges@bidmc.harvard.edu. Gaenna Rogers can be reached with email at grogers1@bidmc.harvard.edu. Please also email other listed on the contact information.

Appointments

For all the other services, appointments are available Monday-Friday from 7 am - 6 pm and must be made in advance with the flow facility. Any appointments outside of scheduled business hours have to be made a week in advance. Monday reservations must be cancelled by close of business (5pm) on Friday prior to the reservation. If a Massachusetts State of Emergency is declared, the core will not be open. Researchers will not be charged for their reservations. Because of the increased demand for sorting and training, please make requests 1-2 weeks in advanced.

Evening appointments are available upon request.

To schedule an appointment for sorting, facility-assisted flow cytometry, training or any other appointment, please call the facility at: Phone: 617-735-4191/4192/4193 or e-mail: jtigges@bidmc.harvard.edu or grogers1@bidmc.harvard.edu. Or anyone else listed on the contact portion of the website.

Sorting

  • Cells should be FILTERED and brought in appropriate tubes:
    • MoFlo: 0.5 - 2 mL Eppendorf Tubes, 5 mL FACS Tubes, and 15 mL Conical Tubes
    • Aria: 5 mL FACS Tubes
  • Cells should be filtered and suspended with concentrations of 20-30 x 10^6/mL and carried in an enclosed container. Please bring collection tubes with media or buffer in the bottom and any necessary antibiotics or antifungals. Sorting is aseptic not sterile. Samples and controls brought from outside of CLS must be transported using secondary containment.
  • The nozzle generates droplets containing cells that the instrument then sorts. Any nozzle switches requires 10 minutes at the beginning and end of your reservation. Nozzle tip default size:
    • MoFlo 7 laser is 70 nm
    • MoFlo 4 laser is 100 nm
    • Aria 5 laser is 85 nm
  • Controls are required for your sort and should include a negative, or unstained control, and single-stained positive control for each fluorochrome you are using. Sorting is aseptic, we are not located in a sterile room, therefore no guarantees of sterility. Your cells will be in a drop with filtered PBS (MoFlo 0.04 nm, Aria 0.2 nm).
  • Sterile collection tubes with media added/extra media. Sorted cells can be collected into:
    • MoFlo: 0.5 - 2 mL Eppendorf Tubes, 5 mL FACS Tubes, and 15mL and 30 mL Conical Tubes
    • Aria: 0.5 - 2 mL Eppendorf Tubes, 5 mL FACS Tubes, and 15 mL Conical Tubes
  • If you are sorting into 96 (or other) well plates make sure they are preloaded with media, and bring extra plates for sorter calibration.

Analyzers

Users are responsible for basic maintenance, including filling the analyzer sheath, emptying the analyzer waste, running a daily clean, and shutting the instrument off if they are the last person on the schedule for the day. Please notify staff if any issues arise with the instrument. If an unexpected issue arises with an instrument the core will notify the scheduled researchers immediately of any delay. We ask for everyone’s patience and understanding in dealing with instrument maintenance.

Booked time is YOUR time, running over appointments-the next booked users have priority. BRING: your own pipets/pipet tips, tubes, gloves, and device for storing your data. Do not leave belongings behind, they will get taken or thrown away. Please properly dispose of trash and unwanted items. Please clean up any spills that were created with provided paper towels, bleach, and water. Anyone left logged in will be charged for excessive time. Anyone using a computer attached to an instrument will be charged. You can only use these analyzers if you have been trained by staff.

Using the Aurora on the weekend does require the user to run QC before running their experiments. The Aurora must have a warm up time of 30 minutes before QC is run.

Data Storage

PC formatted USB, external hard drive, or other storage device should be used to store data. Please take your data right as you finish your experiment. Your data is your responsibility!!! Data dumps will occur to ensure that software is able to run smoothly. Data will never be saved over a year. Any data left longer than a month can be deleted by facility and user charged for time. Data dumps will happen at the end of every month and as needed.