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Flow cytometry

The CHIRI Flow Cytometry facility houses one cell sorter, three analysis platforms and two data analysis computers within its PC2 facilities.

Learn more about CHIRI’s Flow Cytometry shared resource facility below.

General
Cell sorting
Analysers
Data Analysis

General

Open hours

Cell sorting and appointments with CHIRI Flow Cytometry facility staff are available 8AM-6PM Monday to Friday (excluding Curtin-observed public holidays). Trained users can access the analysis platforms 24/7.

Attribution and authorship

All data generated using the shared equipment should acknowledge the CHIRI Flow Cytometry Facility, and where any member of the CHIRI Flow Cytometry Facility has made substantial intellectual contributions to the design of your experiment, they should qualify among the authorship list.

Cell sorting

Single cell sorting

BD FACSJazz cell sorter

BD FACSJazz cell sorter

Details

The BD FACSJazz is equipped with a blue [488nm] and red [635nm] laser and can detect up to six fluorescent colours. It has a fixed 100µm nozzle and, depending on cell type and sample preparation, can sort through 30 million cells per hour.

Sorting

The BD FACSJazz is capable of sorting up to two populations simultaneously into the following collection vessels; single-way sorting into 50ml tubes and cell culture plates (6-, 12-, 24-, 48-, 96- and 384-wells) and two-way sorting into 10-15ml and 5ml tubes.

Instrument specifications | BD FACSJazz Configuration Guide

Analysers

Cell analysers

Attune Classic

Details

The Attune Classic is equipped with two lasers (Blue 488nm / Red 630 nm) and can detect up to six fluorescent channels. Acoustic focusing technology enable high volume, low coincidence single event analysis, of a wide range of particle sizes.

Instrument specifications | Attune Configuration Guide

Instrument performance | Attune Spillover Spread

Canto-II

Details

The BD Canto-II has three lasers (Blue 488nm / Red 633nm / Violet 405nm) and detects up to eight fluorescent channels simultaneously.

Instrument specifications | Canto-II Configuration Guide

Instrument performance | Canto-II Spillover Spread

LSRFortessa

Details

The BD LSRFortessa (Special Order Research Product) is equipped with five excitation lasers (Blue 488nm / Red 640nm / Violet 405nm / UV 355nm / Yellow-Green 561nm) and detects up to 18 fluorescent channels simultaneously.

Instrument specifications | LSRFortessaSORP Configuration Guide

Instrument performance | LSRFortessa Spillover Spread

Data Analysis

Data management

Users are responsible for their own data safekeeping and record keeping. There are automated methods for storing data in multiple locations at the time of sample acquisition and will be covered in the instrument training procedures below.

Data Analysis Suite

The Data Analysis Suite maintains two analysis computers (Mario and Luigi), located on the first floor of Building 305, Room 144. Current staff/student swipe cards are needed to access these platforms.

Analysis software

The analysis computers host multiple software packages, not just for flow cytometry data analysis. For a full list of software available, please see Data Analysis Suite.

  • FlowJoTM(version 10)
  • FlowLogicTM (version 7)
  • Attune Classic Software
  • FCAP Array (cytokine bead array software)
  • GraphPad PRISM (version 7)

Training and bookings Instrument training and booking procedures

Training

Training is a prerequisite to independent access to the CHIRI Flow Cytometry shared resource equipment, and the minimum training requirements are outlined below:

1. Attend a General Flow Cytometry Induction

All users must attend a General Flow Cytometry Induction. Supervisors (or anyone involved in the interpretation of flow cytometry data generated at CHIRI Flow Cytometry Facility) must also attend a General Flow Cytometry Induction. General flow cytometry inductions take approximately three hours and involved two components: theory and practical.

Theory | The theory component covers basic principles of the flow cytometer; including fluidics, optics, electronics and how flow cytometry data is generated. Theory also covers the capabilities and specifications of the flow cytometry analysis and sorting instruments available at CHIRI Flow Cytometry Facility, and an overview of the online booking systems.

Practical | The practical component is laboratory based, and includes correct start-up, cleaning, shut-down, general troubleshooting and data management standard operating procedures. Hands-on instrument training with fluorescent beads is provided to highlight common flow cytometry principles.

2. Follow-up training on instrument of choice

After the general induction and a discussion on the type of flow cytometry application you want to conduct, instrument specific follow-up training is advised. Pilot/preliminary experiments with representative samples and experimental conditions are an ideal opportunity for follow-up training. The follow-up training enables CHIRI Flow Cytometry Facility staff to sign-off user competency, and each have independently have met the minimum requirements for using the shared equipment, know some general troubleshooting procedures, and know the emergency assistance procedures to ensure the longevity of the shared equipment and the quality of data generated. Follow-up training is also an opportunity to set-up application specific and/or cytometer specific settings and templates for future experiments

3. Additional training

Additional training on shared equipment is always encouraged.

Refresher training | If it has been more than six months since you have utilised the flow cytometry platforms or would like to use several flow cytometry platforms, we encourage you to get a refresher training (it takes approximately 30 minutes). Please contact CHIRI Flow Cytometry Facility staff (see ‘Contact us and feedback’ section below).

Additional training | At times additional training and information is necessary, if so please utilise the experienced CHIRI Flow Cytometry Facility staff. However, if the request requires in-depth intellectual input into optimising your flow cytometry application, please refer to our ‘Attribution and authorship’ section above.

Bookings

For cell sorting bookings and flow cytometry appointments, please contact CHIRI Flow Cytometry Facility staff (see ‘Contact us and feedback’ section below).

Access to platform specific online booking calendars is reserved for licensed users only. If you have not received access to the flow cytometry analysers and/or the analysis computers after attending training, please contact CHIRI Flow Cytometry Facility staff (see ‘Contact us and feedback’ section below).

Contact us and feedback Who to contact about flow cytometry

CHIRI’s Flow Cytometry Facility Staff are always interested in ways to improve the equipment and services we offer to support your research endeavours. If you have an enquiry or want to provide feedback, please contact:

CHIRI Facilities Manager

Rob Steuart

Email | r.steuart@curtin.edu.au

Tel | +61 8 9266 7362

Mobile | 0419 192 971

CHIRI Senior Technical Officer (Flow Cytometry)

Jeanne Edmands

Email | Jeanne.lemasurier@curtin.edu.au

Tel | +61 8 9266 7512

Mobile | 0434 987 741