School of Medical Sciences


Flow cytometry and cell sorting facility

What is flow cytometry?


Flow cytometry is the measurement of phenotypic, biochemical or molecular characteristics of cells as they flow past a laser beam creating signals which are collected by a series of detectors. The detectors translate light signals into electronic signals which are then recorded and analysed by computer. Light scatter from cells and fluorescence from the cells themselves or molecules attached to the cells can be measured.

Cell properties such as size, internal complexity, internal and external cell components and DNA content can be measured. Cell sorting takes flow cytometry a step further where populations of cells can be selectively deposited into collection vessels for further culture or analysis.

The Becton Dickinson LSRII flow cytometer has 3 lasers for excitation of fluorochromes: 355nm (UV), 488nm (blue-green), and 633nm (red), and can detect up to 9 colours of emitted fluorescence.

The BD FACSVantage cell sorter has 2 excitation lasers: a 488nm, and a tunable laser capable of emitting UV, blue, green, yellow, and red wavelengths. Up to 5 colours of fluorescence can be detected.

Research using flow cytometry and cell sorting


There are a number of areas where flow cytometry in an invaluable tool for research:

  • Phenotypic analysis of intracellular or extracellular markers
  • Cell cycle analysis
  • Analysis of GFP expression in mammalian cells
  • DNA content determination
  • Intracellular cytokine detection
  • Calcium flux
  • Purification of small populations of cells for PCR
  • Purification of cell populations based on internal or external markers
  • A wide range of other uses

lsr

The facilityLSR

The LSRII Flow Cytometer and FACSVantage Cell Sorter are both located in room 4310 in Building 504 at the FMHS, Park Road, Grafton. Trained staff are available by arrangement to help you run and analyse your samples. LSRII users are able to book time on the machine and run and analyse samples independently. Researchers requiring cells to be sorted must arrange a time with the FACSVantage Operator who will sort your samples for you. The FACSVantage is a less automated machine and requires specialised training.


Costs

Both machines are normally charged out at $108 per hour for internal users (University of Auckland research groups) and $216 per hour for external users. All prices are exclusive of GST.


Booking

A booking sheet for the FACScan is located on the door of room 4310. Users simply fill in their name, contact details and expected time of usage. Actual time used is recorded in a logbook located beside the machine.

FACSVantage users must book time by contacting that facility to arrange a convenient time to discuss requirements prior to cell preparation.

Contacts