Auckland Cytometry

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

Dr Anna Barry, Auckland Cytometry
Dr Anna Barry, Auckland Cytometry

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

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.

Equipment & Services

  • BD Accuri C6 flow cytometer
  • Cytek Northern Lights spectral analyser       
  • Cytek Aurora spectral analyser
  • BD SORP FACS Aria II cell sorter
  • MAGPIX bead assay reader
  • Genomic Cytometry: Single cell RNAseq
Image Component

BD Accuri C6 flow cytometer

2 laser/4 colour analyser
Configurations: 3-blue, 1-red or 2-blue, 2-red

Configuration one: 3-blue, 1-red

Laser
Detector Filter  Common Fluorophores
Blue laser (488nm)
FL1 533/30  FITC, Alexa Fluor488, CFSE, GFP, Cell tracker Green, FDA
Blue laser (488nm) FL2 585/40 PE
Blue laser (488nm) FL3 670 LP PerCP, PE-Cy5, PerCP-Cy5.5, PE-Cy7, PI, 7-AAD
  FL3 610/20 RFP, PI, PE-CF594
Red laser (640nm) FL4 675/25 APC, Alexa Fluor647

Configuration two: 2-blue, 2-red

Laser
Detector Filter  Common Fluorophores
Blue laser (488nm)
FL1 533/30  FITC, Alexa Fluor488, CFSE, GFP, Cell tracker Green, FDA
Blue laser (488nm) FL2 585/40 PE
Red laser (640nm)
FL3 780/60
APC-H7, APC-Cy7
  FL4
675/25
APC, Alexa Fluor647

Example applications

  • Accurate cell counting
  • Cytometric bead assays
  • Real-time functional assays, eg calcium flux (unique to this machine)
  • Automated sampling (24 tube rack or 96 well plate, unique to this machine)
  • 1-4 colour experiments:
    • Simple immunophenotyping

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:

  • 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 (Cytek instruments)
  • Microvesicle studies (Cytek Aurora)

Cytek Northern Lights spectral analyser

  • 2 laser (blue/red)
  • 24 channels (up to 16 colours)

This will be upgraded to include a violet laser. Contact us for the full specifications of the instrument

Cytek Aurora spectral analyser

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

High throughput auto-sampler will be coming soon! Contact us for the full specifications of the instrument.

Cytek Aurora
Cytek Aurora

Cell sorter

BD SORP FACS Aria II cell sorter

  • 4 lasers (blue/red/UV/violet)
  • 17 colour analyser and 4-way cell sorter

Example applications:

  • Multicolour Immunophenotyping (up to 16 colours simultaneously)
    • Cell lines, isolated primary cells
    • Single cell digestions of complex tissue (eg, skin, fat, lymph nodes)
  • 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
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    

MAGPIX Bead assay reader

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

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
    • Cell sorting/viability assessment/sample “clean-up” (Auckland Cytometry)
    • CITEseq/Hash tagging (Auckland Cytometry)

10x Chromium Single Cell 3’ Libraries preparation and Next Generation Sequencing (Auckland Genomics)

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.

Booking: Equipment and services are booked through iLab.

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 dongle-run software package FlowJo by TreeStar which is compatible on both PCs and Macs. FlowJo tutorials can be accessed from their website http://www.flowjo.com/tutorials/. Depending on demand we can lend out these dongles. Please contact us for more information.

Collaborations: 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.

Our people

Director Dr Anna Brooks

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