DHVI Flow Cytometry

The DHVI Research Flow Cytometry Facility serves the analytical and cell sorting needs of the Duke Human Vaccine Institute and researchers throughout the Duke Community. DHVI Flow offers state-of-the-art cytometric support to investigators in basic, developmental, and clinical research.

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).

Download a poster here

Announcements 

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.

 

Here are details on reserving individual instruments:

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 dhviflo@dm.duke.edu.
  • At this point, we are not offering training for new independent users, but are working on a training plan with appropriate social distancing policies. We are, however, 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 Webinars

Tuesday, August 4th - Wednesday, August 5th.

CYTO VIRTUAL 2020, sponsored by the International Society for the Advancement of Cytometry.  Description by Joni S. Moore, Phd, President, ISAC:

We anticipated welcoming you all to my hometown of Philadelphia for CYTO 2020 around now, but biology threw us a curve ball. As scientists, and especially cytometrists, we are all too familiar with having to create new solutions to a never-ending menu of challenges. Although SARS-CoV-2 may keep us from celebrating all the new developments in cytometry in person, we know that you all are still eager to learn about what is happening in cytometry this year and what’s on the horizon.  

So, we’re not letting coronavirus stop us.  We’ve put together a virtual event that will bring much of what we had planned for Philadelphia, as well as some new and timely offerings. Join us on August 4-5 for some cutting-edge programming that will explore the latest developments in flow and image cytometry, and help pave the way for new understandings in basic cellular and molecular mechanisms and human disease.  In light of the COVID-19 pandemic, we will also highlight content related to sharing information on how the cytometry world plays a vital role in bringing meaningful data toward identifying the basic biology of the virus, guiding the way to effective treatments, making faster and more accurate diagnoses, and managing patient care.

Joining us during the conference are 85+ exhibitors from around the world, nearly 150 posters, commercial scientific tutorials, networking opportunities and over 15 hours of live sessions, all of which will be recorded and available on-demand after the conference.  Attend at your convenience!  Registration is here.

 

Wednesday, August 5th, 11:00AM EDT.  T Cell Characterization Combining Advanced Flow Cytometry with Live-Cell Analysis, sponsored by Sartorious.  In this webinar discussion will include how advanced flow cytometry can be used to link T cell function to marker expression and cytokine release, the characterization of bispecific antibodies to reveal mechanistic insights to actions on T cell activation, killing, exhaustion, and memory subpopulations, and how live-cell analysis can complement advanced flow cytometry to provide additional insights to T cell function.  Register here.  Participants can request a certificate of attendance for continuing education purposes.

Thursday, August 6th, 11:00AM EDT.  From B Cells to Yeast - Diverse Approaches to Rapid Discovery of COVID-19 Therapeutic Antibodies, presented by Sony Biotechnology.  Learning Objectives: 1. Gain understanding of flow cytometry, bioinformatics, and synthetic biology techniques that are used for mAb discovery and validation. 2. Understand the end-to-end workflow of screening patient B cells for discovery and stratification of COVID-19 therapeutic antibodies. 3. Learn how yeast surface display is used for accelerated evolution and isolation of high affinity antibody variants against COVID-19.  Participants can request a certificate of attendance for continuing education purposes.  Registration here.

Thursday, August 6th, 1:00PM EDT.  Intro to FlowJo v10.7.  Learn how to use the FlowJo™ workspace, including how to load samples (experimental data), statistics, and gates, create groups and analyses, and generate tabular and graphical layouts. Designed for those new to FlowJo or who would like a refresher overview of the program. Register.

Tuesday, August 11th, 12:00PM EDT.  Advances in Multimodal Informatics: TCR Diversity in Stimulated T Cell Samples, hosted by FlowJo. Analysis of single-cell RNA sequencing data from replicates of sorted T Cell samples; untreated control, and peptide stimulated conditions. Samples were multiplexed using the BD Rhapsody™ Single-Cell Multiplexing Kit. Cells were then sequenced using a targeted panel of 400 genes, 25 surface receptors from BD AbSeq, and T Cell repertoire detected using the BD Rhapsody™ Single-Cell Analysis System VDJ CDR3 protocol. Powerful informatics combined with this cutting edge technology allow us to peer into a diverse compartment of the immune system in high resolution. Register.

Wednesday, August 12th, 1:00PM.  Intro to SeqGeq v1.6, hosted by FlowJo. Single-cell RNA sequencing technologies are rapidly modifying the definitions of extant cell phenotypes. However, the analysis tools used to discover these novel populations remain beyond the reach of most bench scientists. SeqGeq™ is a desktop bioinformatics platform designed from the bench scientist’s perspective; datasets are composed of different types of cells, how do we identify their phenotypes? In this tutorial you will learn how to utilize quality control, normalization, visualization and clustering tools to identify populations of cells in a data set. Once populations have been found, you will learn how to identify differentially expressed genes that define any given population and link those genes to downstream analyses. Register.

Wednesday, August 26th, 11:00AM EDTLearn FlowJo AutoSpill.  FlowJo Product Manager John Quinn, PhD, will be diving into how to use AutoSpill for better compensation.  You will learn how to improve all cytometry experiments, how to make cytometric compensation easier, and improvements and advantages of the AutoSpill approach.  Register.

Thursday, August 20th, 1:00 EDT.  HyperFinder in FlowJo v10.7, hosted by FlowJo. HyperFinder can recapitulate a population, whether it was identified by clustering or a deep manual gating hierarchy, in a specified number of polygon gates. Combined with FlowJo's ability to export to BD FACSDiva™, this can be incredibly valuable for sorting more complex populations than was previously possible. In this webinar we will demonstrate how to get HyperFinder from FlowJo Exchange, install it, use it, and export the resulting gates to BD FACSDiva. Register.

Friday September 18th, 1:00PM EDT.  Open Flow (a collaboration between the Francis Crick Institute and the Memorial Sloan Kettering Cancer Center) will host their fourth online flow cytometry educational session and will focus on troubleshooting and understanding common mistakes in compensation utilizing BD FACSDiva.  All are welcome.  Register here!

(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 the session can be found on their YouTube.)

 

 

 

 

 

Facility Leadership

  • 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