School of Medicine:

Established in 1930, Duke University School of Medicine is the youngest of the nation’s top medical schools. Ranked tenth among its peers, the School takes pride in being an inclusive community of outstanding learners, investigators, clinicians, and staff where traditional barriers are low, interdisciplinary collaboration is embraced, and great ideas accelerate translation of fundamental scientific discoveries to improve humanhealth locally and around the globe.

Comprised of 2,400 faculty physicians and researchers, the Duke University School of Medicine along with the Duke University School of Nursing and Duke University Health System create Duke Health. Duke Health is a world-class health care network. Founded in 1998 to provide efficient, responsive care, the health system offers a full network of health services and encompasses Duke University Hospital, Duke Regional Hospital, Duke Raleigh Hospital, Duke Primary Care, Private Diagnostic Clinic, Duke Home and Hospice, Duke Health and Wellness, and multiple affiliations.

This position is grant-funded.

Job Description

The Burt laboratory is seeking an inquisitive, motivated, and organized individual to join the research team in the Department of Pediatrics (Division of Neonatology and the Children’s Health and Discovery Initiative) and the Flow Cytometry Shared Resource in the Duke Cancer Institute.

This position is located in the Chesterfield Building location in Durham, N.C.

This individual will have duties both as the Burt lab manager and lead research analyst, and as a flow sorter operator for the Duke Flow Cytometry Shared Resource in the Duke Cancer Institute. The individual will manage operational aspects of the Burt lab, and also engage in personal experimental work with projects focused on understanding immune system development and function in the human newborn infant, particularly in the setting of infection and vaccination. In their capacity as flow sorter operator, the individual will work with clients to sort-purify cells from a variety of biological sources and experimental setups.

Applicants with experience in common cell-based immunology assays, flow cytometry phenotypic analysis and sorting, blood and tissue processing, and basic molecular biology techniques who also have prior experience in lab management are strongly preferred.

Occupational Summary

Conduct a variety of complex and independent investigations in the planning, development and implementation of experimental procedures and support of neonatal/pediatric research activities at Duke University. Research activities conducted by this individual will center on in vitro assessment of biological samples from human blood and tissue samples. Additionally, this individual will perform sample preparation, staining, and FACS sorting for clients engaging the Duke Flow Cytometry Shared Resources’ services for cell sorting. This position involves daily and periodic maintenance and upkeep of a dedicated FACS sorter located in the Chesterfield Building.

Work Performed

Conduct independent investigations in the planning, development and implementation of original experimental procedures, including physical, chemical, biological and other research laboratory tests, to obtain data for research purposes.

Compile, analyze, document procedures and results, and enter data into relevant research databases and laboratory documents; draw tentative conclusions from experimental results.

Review laboratory methods manual, scientific journals, abstracts and other literature for information applicable to research experiments.

Confer with principal investigator to review work assignments and develop plans for research experiments; make modifications to existing laboratory procedures and techniques to meet the needs of the particular equipment as required.

Prepare tissue and specimens for complex procedures, and devise experimental techniques for tests and analyses. These projects involve extensive knowledge of tissue culture and flow cytometry. Focus of work will be preparation and assessment of tissues and cellular samples from human pregnancy, including maternal, paternal, and umbilical cord blood, and biological samples of lymphoid and gut tissue from children and adults (e.g., surgical samples of thymus, tonsil, lymph node and intestine). Predominant method of analysis will be based in flow cytometry methods.

Prepare and enrich cell populations derived from human umbilical cord blood and human adult peripheral blood using immunomagnetic separations, flow cytometry, or other selection techniques. This includes regular Ficoll preparation of peripheral blood mononuclear cells (PBMCs) from a variety of human clinical samples. Perform cell culture, mixed lymphocyte reactions, cell suppression assays and assist with characterization of the biological activities of cell populations. Other analyses may include transfection of cells for gene editing with CRISR-Cas9 reagents, PCR, RT-PCR, RNA sequencing, ChIP sequencing, DNA methylation pyrosequencing, ELISA (multiplex ELISA), cytokine bead array, histological staining, immunohistochemistry or immunofluorescence, image analysis and other methods as appropriate. Document all experimental procedures and results in format suitable for maintenance of intellectual property and for regulatory audits.

Evaluate and interpret collected data and prepare reports and analyses setting forth progress, adverse trends and appropriate recommendations or conclusions. Confer with principal investigator in developing plans for research projects and to discuss the interpretation of results and the preparation of scientific manuscripts for publication.

Set up instruments, material and apparatus and operate standard laboratory equipment required for specific tests. Operate various electronic devices in the collection, tabulation and calculation of data. Obtain, prepare, mount and stain tissue, blood and other samples for microscopic examinations. Prepare culture media, chemicals, reagents and solutions.

Review journals, abstracts and scientific literature to keep abreast of new developments and to obtain information regarding previous experiments to aid in the design and development of original procedures and techniques. Assist in the development of new research procedures and techniques.

Supervise, train and instruct residents, interns, students or lower level personnel in laboratory techniques and procedures. Review work of lower level laboratory personnel and assist in the training of new personnel (staff, students, and post-doctoral scholars), including training in aspects of lab work safety. Maintain human research ethics training, and other laboratory training records, for self and other lab members. Interface professionally and collegially with other members of the Burt lab and collaborating labs. Careful and routine maintenance of biological sample databases, keep up to date intake and use of specific samples for various projects in the lab.

Maintain sufficient inventory of materials, supplies and equipment for performance of duties, including serving as the primary agent for ordering through the online purchasing portal. Serve as an interface with vendor sales representative. Serve as a first contact, along with the PI, for equipment alarms and emergency management. Help maintain compliance with regulatory guidelines including Duke Biological and Chemical Safety. Complete routine laboratory cleaning and maintenance.

Perform other related duties incidental to the work described herein.

Skills:

Experience with tissue culture and flow cytometry is absolutely required. A minimum of 3 years (greater than 5 years preferred) hands-on experience as follows:

• Isolation and cell culture (with excellent sterile technique) of primary cells from human or animal tissues and/or human blood primary cells
• Preparation of cells for flow cytometry, including staining with fluorophore-conjugated monoclonal antibodies (mAbs) and Live/Dead staining.
• Immunomagnetic cell sorting for sample purification for direct culture applications or as enrichment prior to FACS sorting
• Strongly prefer specific experience with stimulation, fixation and permeabilization for intracellular cytokine staining and/or phospho-flow, as well as assessment of cell proliferation by traceable dye dilution (e.g., CFSE, cell-tracker dyes)
• Titration of antibodies for optimal staining concentrations
• Construction and assessment of multiparameter flow-cytometry panels, including assessment and trouble-shooting of compensation controls, familiarity with fluorophore characteristics and knowledge of compatibility of fluorophores in multicolor panels
• Experience and facility with data analysis using standard flow cytometry analysis software (e.g., Flow-jo, Diva) is required
• Experience and facility are required with data analysis of processed flow cytometry data, including graphing, figure preparation, and presentation using at least one of the following analysis programs, and willingness to learn others: Microsoft Excel, Graph Pad Prism, JMP, or R/bioconductor

In addition to basic skills described above, the role of Flow Cytometry Core Lab FACS operator requires additional skills related to flow cytometry. A minimum of 3 years (>5 years preferred) hands-on experience as follows:

• Hands on set-up, operation, trouble-shooting, and maintenance of analytic flow cytometers and flow cytometry-based cell sorters, including daily and periodic quality control and quality assurance testing.
• Must be able to diagnose and fix basic problems during routine flow cytometry analysis and sorting (e.g., clogs, inappropriate compensation), and recognize issues requiring professional service (e.g., laser or fluidics malfunctions).
• Must be able to adapt to a wide variety of cell types, sizes, and physiochemical properties and tailor staining and flow cytometry regimens to meet individual needs of clients.
• Manage data acquisition, as well as data entry and storage for Flow Cytometry Core clients.

Experience with specialized immunological assays. Work experience in a cellular immunology laboratory and hands-on experience with routine immunological assays is required (minimum 3 years, preference for greater than 5 years). The following specific techniques are strongly preferred:

• In vitro mixed lymphocyte reactions (MLR) and anti-CD3 mAb stimulation to assess T cell activation and proliferation
• Intracellular flow cytometry for cytokine production
• ELISA and bead-array based techniques for assessment of cytokine secretion

Management and organizational skills. Given the managerial nature of the position, applicants require a minimum of 3 years (greater than 5 years preferred) in a management position as either a single lab manager, manager for a collaborative group of labs, or core lab manager.

• Project and experiment planning and management for multiple simultaneous projects and clients
• Setting and meeting benchmarks in a timely fashion
• Assessment, inventory management, and record keeping of lab supply stock, anticipation of needs for upcoming activity, ordering and proper storage of supplies (for both Burt lab and Flow Cytometry Core lab)
• Excellent interpersonal and communication skills, both written and oral
• Problem solving
• Attention to detail

Experience with sterile techniques/methods A minimum of 3 years (5 years strongly preferred) experience with cell/tissue culture or primary cells and/or cell lines requiring sterile technique is required. Experience working in a BSL2 environment (esp. primary human cells) is strongly preferred, or willingness to learn Universal Laboratory Precautions for compliance with laboratory safety and infection control procedures and practices.

Minimal require biochemistry and molecular biology experience:

• Small and larger scale nucleic acid and protein purification, including meticulous clean technique and operating procedures to avoid contamination
• Planning and execution of basic DNA PCR and RT-PCR experiments, including troubleshooting
• DNA and protein gel electrophoresis and image capture
• Procedures involved in cell transfection, preparation of nucleic acid for DNA sequencing, RNA sequencing (RNA-seq; including cDNA library preparation) and/or microarray analysis are not required, but strongly preferred.

Minimum Qualifications

Education

Work requires a bachelor's degree in botany, biology, zoology, psychology or other directly related scientific field.

Experience

Work requires four years of research experience. A related master's OR AN EQUIVALENT COMBINATION OF RELEVANT EDUCATION AND/OR EXPERIENCE degree may offset required years of experienceon a 1:1 basis, e.g., a two year master's degree in lieu of two years of experience.

Duke is an Affirmative Action/Equal Opportunity Employer committed to providing employment opportunity without regard to an individual's age, color, disability, gender, gender expression, gender identity, genetic information, national origin, race, religion, sex, sexual orientation, or veteran status.

Duke aspires to create a community built on collaboration, innovation, creativity, and belonging. Our collective success depends on the robust exchange of ideas—an exchange that is best when the rich diversity of our perspectives, backgrounds, and experiences flourishes. To achieve this exchange, it is essential that all members of the community feel secure and welcome, that the contributions of all individuals are respected, and that all voices are heard. All members of our community have a responsibility to uphold these values.

Essential Physical Job Functions: Certain jobs at Duke University and Duke University Health System may include essentialjob functions that require specific physical and/or mental abilities. Additional information and provision for requests for reasonable accommodation will be provided by each hiring department.