Flow Cytometry Core Facility (FACs Core)

Overview:

The Flow Cytometry Core Facility (FACs Core) at the Translational Research Institute (TRI) offers access to high-end FACS analysis and state-of-the-art cell sorting capabilities, actively providing scientists with adaptive setups and dedicated support. Flow cytometer/fluorescence activated cell sorting (FACS) represents fast and reliable methodology for characterizing and isolating cell populations and subpopulations, as well as to quantify soluble particles. It is based on a single cell suspension (or other soluble particles), which individually passes through a focused laser beam and the characteristics scattered and fluorescent light generated are detected separately. Fluorescence dye-labelled antibodies are generally used to stain the cells, which specifically recognize and bind structures on the cells. By suitable selection and composition of the individual markers with different fluorescent dyes, several dozens of subpopulations or soluble particles can be distinguished in one panel of a sample material.

Due to the continuous technical development and the use of novel fluorescent dyes, the field of application of flow cytometry has steadily expanded in recent years, especially in single cell analysis (e.g. genome and transcriptome analysis), identification, phenotypic and functional characterization of rare cell population, cell sorting and quantification of soluble mediators.

Core Services and Technologies:

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The FACs Core facility provides a wide range of services available to researchers and clinicians within HMC as well as other institutes within Qatar who are interested:

• Immune phenotyping of cell population in a clinical sample.
• Identification and characterization of rare cell population/s in a clinical sample.
• Intracellular cytokines and chemokines analysis.
• Apoptosis (Annexin, PI, Cytochrome C, MitoStatus etc.)
• Cell proliferation
• Cell Cycle
• Autophagy
• Intracellular calcium influx
• Cytokine bead array
• Cell sorting
• Index cell sorting for sequencing
• Consultation on experimental design, sample preparation, data acquisition, analysis and interpretation

User information:

The following procedures must be followed for all users:

• To avail the services of the FACs core, the interested researchers must submit the project details to the core facility lead. Once the project details have been reviewed by the core it will be forwarded to TRI management for scoring and approval.
• When you have prepared your test samples according to the recommendations made during the initial consultation with the core facility, we will introduce you to the instruments making a basic acquisition setup with your samples or beads. An introductory training and brief formal orientation are mandatory for all users before starting to use the facility. We do not permit access to the instruments for users who are not trained directly by Core staff. After acquisition of your data, we will show you the possibilities of data analysis located at the core facility.
• Second, it is imperative that you carefully plan your experiments in advance. The Flow core is always available to consult on protocols and procedures prior to implementation of your assay. Any non-routine multicolor panel experiment you wish to do should have a pilot experiment done beforehand. It is imperative that you discuss any complex multicolor experiment with the core facility team prior to planning and/or ordering supplies to avoid any problems or misunderstandings.

Important information - Cell sorters are run ONLY by core facility employees and all experiments must be discussed beforehand and scheduled with a core facility employee in advance.

Instruments and technology:

The instruments and technology of the FACs core includes:

• Cytek^TM Aurora
  This is a spectral flow cytometer with up to five lasers (Ultraviolet 355nm, Violet 405nm, Blue 488nm, Yellow Green 561nm, Red 640nm). It can measure up to 64 fluorescence parameters (UV1-UV16, V1-V16, B1-B14, YG1-YG10, R1-R8) in a single tube/sample. This system delivers high quality data where rare and dim populations are easily resolved, and it enables the use of a wide array of new fluorochrome combinations without reconfiguring the system for each application. It has the SpectroFlo® software that offers an intuitive workflow from quality control to data analysis with technology-enabling tools that simplify running any application.

• BD FACSymphony^TM A5
  The BD FACSymphony™ A5 cell analyzer enables the simultaneous detection of up to 30 parameters in a single tube/sample. It comprises 5 lasers (Ultraviolet 355nm, Violet 405nm, Blue 488nm, Yellow Green 561nm, Red 640nm).

• BD LSRFortessa™
  This cell analyzer has 5 lasers (Ultraviolet 355nm, Violet 405nm, Blue 488nm, Yellow Green 561nm, Red 640nm) and enables the simultaneous detection of up to 20 parameters in a single tube/sample.

• BD FACSAria ^TM III instrument
  This instrument is a cell sorter with highly sensitive analysis performance. It can detect up to 20 parameters and comprises 5 lasers (Ultraviolet 355nm, Violet 405nm, Blue 488nm, Yellow Green 561nm, Red 640nm). It has the following analytical characteristics:
  4 ways sorting from a single sample
  Bulk and single cell sorting
  Index sort and nozzle size: 70µm, 85µm and 100µm

Booking of instruments

All instruments (BD LSR Fortessa, BD Symphony A5, Cytek Aurora and BD FACS Aria III cell sorter) can only be booked by the core facility staff. Please ask in advance in case you need to use one of these instruments. Booking dates can be requested from Sunday to Thursday between 7:30 am (start time) to 3pm (end time). Bookings that do not fall into this time range have to be specially arranged with the staff.

NO-SHOW POLICY. Users should give at least 24 hours’ notice if they need to cancel an appointment. If a user is late for an appointment by more than one hour, then it is their responsibility to determine whether the required work can still be done.

Contact Details:

Translational Research Institute (TRI)
Hamad Bin Khalifa Medical City, Building 320, 2^nd Floor, Room No. 2-3-4
Hamad Medical Corporation (HMC), P.O. Box 3050, Doha, Qatar

Dr. Maysaloun Merhi (Head, Flow Cytometry Core Facility)
Translational Research Institute (TRI), Academic Health System (AHS)
MMerhi@hamad.qa
+97444397805

Dr. Fareed Ahmad (Co-Head, Flow Cytometry Core Facility)
Translational Research Institute (TRI), Academic Health System (AHS)
FAhmad8@hamad.qa
+97444395238

Collaborating Institutes:

Dermatology Institute, NCCCR, Qatar Metabolic Institute.

Our Research:

1. Ansari AW, Ahmad F, Raheed T, Jochebeth A, Mateo JMP et al. Azithromycin downregulates ICOS (CD278) and OX40 (CD134) expression and mTOR activity of TCR-activated T cells to inhibit proliferation. Int Immunopharmacol. 2023 Nov;124(Pt A):110831. doi: 10.1016/j.intimp.2023.110831.
2. Mestiri S, Merhi M, Inchakalody VP, Taib N, Smatti MK et al. Persistence of spike-specific immune responses in BNT162b2-vaccinated donors and generation of rapid ex-vivo T cells expansion protocol for adoptive immunotherapy: A pilot study. Front Immunol. 2023 Feb 2;14:1061255. doi: 10.3389/fimmu.2023.1061255.
3. Kuttikrishnan S, Masoodi T, Ahmad F, Sher G, Prabhu KS et al. In vitro evaluation of Neosetophomone B inducing apoptosis in cutaneous T cell lymphoma by targeting the FOXM1 signaling pathway. J Dermatol Sci. 2023 Nov;112(2):83-91. doi: 10.1016/j.jdermsci.2023.10.001
4. Abdulrahman N, Leo R, Boumenar HA, Ahmad F, Mateo JM et al. Embelin inhibits viability of cutaneous T cell lymphoma cell lines HuT78 and H9 by targeting inhibitors of apoptosis. Leuk Lymphoma. 2023 Dec;64(14):2236-2248. doi: 10.1080/10428194.2023.2256909
5. Kuttikrishnan S, Ahmad F, Mateo JM, Prabhu KS, El-Elimat T et al. Neosetophomone B induces apoptosis in multiple myeloma cells via targeting of AKT/SKP2 signaling pathway. Cell Biol Int.2024 Feb;48(2):190-200. doi: 10.1002/cbin.12101​