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
The facility
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
