The Flow Cytometry Facility instruments are capable of performing BSL-1 through BSL-3 live cell sorting, phenotypic acquisition, DNA cell cycle analysis, and intracellular marker analysis. Analytical and sorting capabilities up to 18 simultaneous parameters enable researchers to define subpopulations based on cell surface morphology as well as size and complexity. We have three cell sorters (2 BD FACS Arias and 1 BD Influx) that are operated by facility staff. We have four cell analyzers (2 BD LSRII, 1 BD Fortessa and 1 Luminex Muse) available 24/7 for trained users. Instruments are located in the RP105, MSRBII, and GHRB (RBL at Duke).
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In response to COVID-19, we are operating with a limited set of services offered by the DHVI Flow Facility.
Under our "Phase III" plan, we have implemented an Enhanced Infection Control Policy, as detailed below:
a. All staff and users to wear masks at all times. It is the user’s responsibility to obtain a mask from their PI/supervisor.
b. Hand-washing: Staff and users to wash hands at arrival and departure from facility labs.
c. Users and staff to wipe down computers (instrument controllers and work stations) and instrument switches (Power, Hi/Low, etc.) with disinfectant wipes (Chlorox disinfectant wipes or paper towels sprayed with 70% ethanol) at beginning and end of each session.
d. Facility staff to wipe down door knobs, sinks, and light switches with disinfectant wipes (Chlorox disinfectant wipes or paper towels sprayed with 70% ethanol) at beginning and end of each day, and at regular intervals (every ~2 hours) during periods of heavy instrument usage.
e. Only one user in a room at a time [the three rooms in RP105 are the main lab (L01/F01), L02 lab, and anteroom]. At most, one user can use the L02 and another the L01 or F01. Because of their proximity, the L01 and F01 cannot be used by different users at the same time – this policy will be enforced through the CoreResearch reservation calendar (see below. Similarly, only one person to be in the anteroom at any given time.
f. When sorting, only one researcher permitted in a room along with the facility staff. Zoom meetings may also be scheduled if a researcher would like to view and discuss their sort data remotely with the facility operator.
Here are details on reserving individual instruments:
A01: calendar is open on CoreResearch
A02: calendar is open on CoreResearch
L01: calendar is open on CoreResearch
L02: calendar is open on CoreResearch
F01: calendar blacked out, but instrument is available by contacting facility staff. To schedule time on the F01, please email the flow facility at DHVIflo@dm.duke.edu with the desired date/time of your reservation and we will open the time up for you. Facility staff will black out the calendar for the L01 for the scheduled time – the goal here is that only one person occupies the L01/F01 space at any given time.
M01: calendar is blacked out, but instrument is available by contacting facility staff. Note that this instrument has been moved to MSRB2 4116. To schedule time on the M01, please email the flow facility at DHVIflo@dm.duke.edu with the desired date/time of your reservation.
- Staff will be on-site only as needed for facility services or maintenance. If you are the first user of the day, please be prepared to run CST. If you are the last user of the day, please shut down the instrument. If you run into issues, please call 684-4130 or email email@example.com.
- Analyzer training has now resumed. We are also still providing facility orientation for new users who wish to perform sorting in MSRB2 (A02) or GHRB (A01).
- If your lab has been approved by the School of Medicine to open, you should have badge access to RP1.
Upcoming and Recorded Webinars
TOPIC: COMPENSATION IN DIVA. Recorded Webinars: Open Flow (a collaboration between the Francis Crick Institute and the Memorial Sloan Kettering Cancer Center). Open Flow's third session, Flourescence Compensation in Diva, reviewed the principles behind compensation and how it is carried out on a BD Fortessa analyzer, using a live-run with a four color experiment. A recording of their third session can be found on their YouTube.)
Also, Open Flow held their fourth session on Friday September 18th, 2020. This session, Multicolor Troubleshooting in Diva, focused on rules of compensation and how our data can suffer when we do not follow them. A recording of this fourth session can be found on their YouTube.
***DHVI FLow Cytometry Shared Resource 2020 FlowJo Training Workshop, Day 1, Basic: Recording posted here*** (Passcode is g7=yETgZ). Topics covered will include loading data, accessing metadata, cleaning data through exclusion gates, compensation, keywords, creating groups, creating gates, applying statistics, working the layout and table editors, and more.
***DHVI FLow Cytometry Shared Resource 2020 FlowJo Training Workshop, Day 2, Advanced: Recording posted here*** (Passcode is 1#AHfDKF) Topics covered include advanced applications in FlowJo, including plugins and how to install them, data pre-processing features (such as DownSample, StainIndex, and concatenation), dimensionality reduction techniques (such as tSNE and UMAP), and clustering tools.
- Director: Derek Cain, PhD
- Flow Engineers: Steven Slater ASCP(SCYM) and Evan Trudeau ASCP(SCYM)
- Research Technicians: Patti McDermott ASCP(SCYM) and Aria Arus-Altuz
- Scientific Advisors: Gregory Sempowski, PhD and M. Anthony Moody, MD