Flow Cytometry

Overview

ARAFlowCore provides world-class flow cytometry services to scientists from Monash University, The Burnet Institute, The Baker Institute, other members of the Alfred Research Alliance and the broader academic and commercial scientific community.

ARAFlowCore is one of the top flow cytometry platforms in Australia with dedicated laboratories housing three high speed cell sorters in PC2 compliant laboratories, six analysers, and an imaging cytometer. Features of the platform include spectral analysers and cell sorter with capacity to visualise 40+ fluorescent parameters on a single cell, and a six-laser cell sorter which can detect fluorochromes from ultraviolet through to infrared wavelengths allowing unrivalled flexibility for facility users.

The platform’s experienced staff operate the cell sorters and provide training and assistance for the analysers, along with expertise in experimental design.

ARAFlowCore is located across level 2 of Monash's Australian Centre for Blood Diseases (ACBD), and level 6 Burnet tower and is a node of the wider FlowCore facility at Monash University.

Expertise

Many areas of biomedical science utilise flow cytometry, including immunology and pathology, medicine, neuroscience, infectious disease, and microbiology.

The platform offers cell sorting services where a live pure population of interest is collected for downstream applications using animal or patient samples. Some examples of facility use includes:

• Immunophenotyping – pure cell populations from a complex cellular environment such as tissue or blood can be characterised and enumerated.
• Studying disease and pathologies – e.g. brain tissue from stroke models can be analysed for microglia populations.
• Supporting clinical studies – immune profiles of patient blood samples are examined from patients with COVID or recovering from COVID.
• Efficiency of crispr/cas9 editing
• Small particle detection such as extracellular vesicles and platelets

Purified cell populations can also be further studied in:

• Single cell RNA sequencing to sort single cells, obtaining protein expression data and comparing that with RNA message in the same single cell.
• Gene expression studies in pure immune cell populations prior and post treatment for diseases such as multiple sclerosis.
• Cell culture, gene editing, and functional assays.

ARAFlowCore also uses the emerging technology of spectral cytometry which greatly enhances the amount of information able to be derived from one sample. It collects and processes fluorescent information differently giving researchers enhanced flexibility in accessing cellular information.

The platform is experienced in working with industry collaborations and is ISO9001 accredited.

Equipment

Cell Sorters

Cell sorting is the process of selecting and separating populations of interest from a heterogeneous sample. Each sort is set up and supervised by a dedicated ARAFlowCore staff member, per the researcher’s requirements.

• BD FACSAria Fusion
  • 6 lasers; 18 fluorescent channels; sterile sorting; 3 nozzles (70um, 85um, 100um)
• BD Influx
  • 4 lasers; 15 fluorescent channels; sterile sorting; 4 nozzles (70um, 86um, 100um, 130um)
• Cytek Aurora Cell Sorter
  • 5 lasers; 64 fluorescent channels; sterile sorting; 4 nozzles (70um, 85um, 100um, 130um)

Analysers

Users are trained to run their own samples on the analysers. Each analyser is capable of manual acquisition, with some capable of high-throughput sampling (HTS).

• Beckman Coulter CytoFLEX
  • 3 lasers, 13 fluorescent channels, 5ml sample tubes, HTS with 96 well plates
• BD LSRII
  • 4 lasers, 16 fluorescent channels, 5ml sample tubes
• BD LSRFortessa
  • 4 lasers, 16 fluorescent channels, 5ml sample tubes, HTS with 96-384 well plates
• BD LSRFortessa X-20
  • 5 lasers, 18 fluorescent channels, 5ml sample tubes, HTS with 96-384 well plates
• Cytek Aurora
  • 5 lasers, 64 fluorescent channels, 5ml sample tubes
• Cytek Northern Lights
  • 3 lasers, 38 fluorescent channels, 5ml sample tubes, HTS with 96-384 well plates
• Amnis ImageStreamX MkII
  • 4 lasers, 10 fluorescent channels, 2 cameras, 1.5ml microcentrifuge sample tubes

Analysis computer

FlowJo software installed and free to use for registered users.

Access and Contact

Completion of a training and induction process is required prior to using the facility and can be arranged via the contact below.

The facility is available to all researchers in academia and industry and bookings are made via the iLabs booking system. For enquiries or more information contact:

Melanie Le Page
Flowcore@ARA Platform Manager
Monash University
melanie.lepage@monash.edu or
ara.flowcore@monash.edu

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