Auckland Cytometry

Use our technical expertise, training and access to state-of-the-art technologies in flow cytometry.

Dr Anna Brooks, Director of Auckland Cytometry
Dr Anna Brooks, Director of Auckland Cytometry

Auckland Cytometry is a flow cytometry core facility which provides researchers with technical expertise, training and access to state-of-the-art technologies and applications in flow cytometry. 

We can support your research by offering access to:

  • training to use the analysers
  • trained staff to assist with analysis or cell sorting
  • consultation or collaborations to assist with experimental design and data analyses
  • data analysis Software

Flow cytometers are able to analyse several thousand cells (particles) every second, in "real time," and specialised cell sorting cytometers can actively separate and isolate cells (particles) having specified properties. Flow cytometry, typically using fluorescent probes which bind to specific cell associated molecules, allows measurements of various molecular characteristics of individual cells (or particles) suspended in a fluid stream. The use of flow cytometry can be divided into two broad categories, analysis and cell sorting.

Instruments:

  • Cytek Northern Lights 3 Lasers spectral analyser with plate loader  
  • Cytek Aurora 3 Lasers spectral analyser
  • Cytek Aurora 5 Lasers spectral analyser
  • BD SORP FACS Aria II cell sorter
  • MAGPIX bead assay reader

1. Spectral Analysers

Spectral cytometry is an exciting new technology we are proud to host at the facility. As opposed to conventional detection, spectral cytometers lack individual detectors for the measurement of a dedicated fluorophore. Rather, a “spectral signature” is registered for each particle or cell that is interrogated, and this spectral signature is deconvoluted in order to extract intensity data for each dye. This means that we can use many more dyes in combinations not possible on conventional instruments.

Example applications on spectral analysers:

  • Multicolour Immunophenotyping

                    - Cell lines, isolated primary cells

  • Fluorescent protein detection (eg eGFP) – transduction efficiency
  • Functional assays (ie live/dead /cell proliferation, cytokine secretion, receptor regulation)
  • Multicolour phenotyping (at least 26 colours on the Cytek Aurora)
  • Autofluorescent cell studies -primary tissue digests/cultured cell lines
  • Microvesicle studies

1.1 Cytek Northern Lights and Aurora spectral analyser

Both instruments have:

  • 3 lasers (blue/red/violet)
  • 38 channels (up to 28 colours)

In addition to acquisition from tubes, the Cytek Northern Lights is compatible with 96-deep is 96-well plate (V, U and flat bottom) offering more versatility when running your samples at high-throughput.

1.2 Cytek Aurora spectral analyser:

  • 5 lasers (blue/red/yellow-green/violet)
  • 50 channels (at least 26 colours)
Cytek Aurora
Cytek Aurora

2. Cell sorter

BD SORP FACS Aria II cell sorter

  • 4 lasers (blue/red/UV/violet)

Cell sorting flow cytometer capable of sorting up to 4- ways simultaneously into tubes, single and multiple cell deposition into 6 to 384 well plates. The Current laser configuration enables the simultaneous detection of up to 17 colours.

Example applications:

  • Single cell deposition (sorting) - cloning from single cells
  • Identification and isolation of transduced (fluorescent) cells
    • Molecular characterisation/functional assays
  • Enriching cells of interest/depleting cells/debris
BD SORP FACS Aria II cell sorter
BD SORP FACS Aria II cell sorter
Detectors   Band Pass   Dichroic Filter Fluorophores        
Blue Laser (488nm)
E 530/30
  505LP FITC Alexa-488 CFSE GFP BB515
  D 575/25
  550LP PE        
  C 610/20
  600LP PE-CF594 ECD PE-TR PI  
  B 695/40
  635LP PerCP-Cy5.5 Pe-Cy5 PerCP-710 PerCP BB700
  A 780/60
  755LP PE-Cy7        
Red laser (640nm) C 660/20     APC Alexa-647      
  B 710/50   685LP Alexa-700        
  A 780/60   755LP APC-Cy7 APC/Fire750      
UV Laser (355nm) C 450/50     C DAPI      
  B 530/30   505LP B BUV395      
  A 780/60   635LP A BUV737 BUV805    
Violet laser (405nm) F 450/50     BV421 CellTrace Violet SB436    
  E 510/50   502LP BV510 BV480      
  D 610/20   600LP BV605 SB600      
  C 660/20
  630LP BV650 SB645      
  B 710/50   685LP BV711 SB702      
  A 780/60   750LP BV785 BV786 SB780    

3. Bead assay reader

MAGPIX 

Highly specialised magnetic-based system that reads multiplexed bead assays in biological samples.

Multiplexing allows multiple biological target analytes to be simultaneously examined and quantified in a single sample.

Bead assay reader
Bead assay reader

Policies, Services and Guidelines

1. How We Can Help You

  • Project Design/method development: we are available for consultation or collaborations to assist with experimental design and data analyses.
  • Training: Researchers can be trained to use the machines themselves or pay to have an operator assist. Cell sorting will generally require an experienced operator to assist, although this can also be trained if the researcher will be performing this regularly.

2. Booking

Equipment and services are booked through iLab.

3. Contact

cytometry@auckland.ac.nz

4. Services and Collaborations

  • We are available to assist you with your project, please Contact us to discuss it. Costs, regents, technical service and sample requirement information will be supplied once we understand your needs and project.
  • We are always keen for new collaborations – especially multicolour panel development, so please get in touch if this is something that would advance your research.
  • We are also always keen to try new techniques and help develop assays.

5. Data Storage and Analysis

Storage of data generated on any of our machines is the responsibility of the user. You will be required to take your data with you following completion of your run. We are not responsible for ensuring your data is backed up. Further instructions will be provided depending on the machine you will be using. Google Drive is an efficient way of transporting your data, as all machines will have internet access.

Data generated from all the instruments is the responsibility of the investigator, as a separate software package is often required. Data analysis is the responsibility of the investigators although training, and assistance can be provided. We use the software package FCS Express, which is compatible on both PCs and Macs. FCS Express tutorials can be accessed from their website here. Please contact us for more information.

6. Acknowledgments

If you are using the Core Facility, please acknowledge the Core facility in all publications and grant applications where equipment use and/or our services or/and expertise were used. One example of an acknowledgement used in publications is: “We acknowledge the Auckland Cytometry Flow Cytometry ShaRE Core for…”

If a staff of the ShaRE team has worked closely and helped you with your project, it is a good habit acknowledge this including their name in the acknowledgement section.

Also, please send us your newly accepted papers when contain data produced at the Core Facility. We enjoy seeing the work that the costumers are generating here!

7. Genomic Cytometry techniques

Single cell RNAseq

  • In collaboration with Auckland Genomics, we can offer access to:
    Consultation or collaborations to assist with cell preparation for single cell applications
    o Cell sorting/viability assessment/sample “clean-up” (Auckland Cytometry)
    o CITEseq/Hash tagging (Auckland Cytometry)
    o 10x Chromium Single Cell 3’ Libraries preparation and Next Generation Sequencing (Auckland Genomics)

Our People

Director
Dr Anna Brooks

Technologist
Thaize Chometon

Assistant Technologist
Tanvi Damani

Dr Anna Brooks at work in the Auckland Cytometry lab
Dr Anna Brooks at work in the Auckland Cytometry lab