Location
Flow Cytometry
Open to staff and students, this core facility promotes the use of flow cytometry in a wide variety of applications. We offer use of two multiuser benchtop flow cytometers.
Flow cytometry basics
In the technique of flow cytometry a population of cells or other particles is passed in single file in a focused stream of flowing fluid through a laser illumination source. Information about the cell or particle may be deduced from the scattered laser light in the direction of the laser path (Forward Scatter) and at ninety degrees to the incident laser light (Side Scatter). Additional information about cell phenotype and function may be gathered by an analysis of light emitted at specific frequencies from excited fluorochromes added to the cells. The flow cytometer provides a platform for the rapid collection of data generated in this way, and this information can be collated into readily decipherable graphical representations.
Flow cytometry is commonly used to investigate:
- Cell immunophenotype using fluorochrome labelled monoclonal antibodies
- The cell cycle
- Cell proliferation
- Cell apoptosis / cell death and viability
- Functional cell assays such as Calcium Release, Phagocytosis, Reactive Oxygen Species Formation and Cytoplasmic Enzyme Reactions
- Intracellular protein phosphorylation determination in cell signalling networks
- Multiple cytokine determination in a single small volume sample
- Cytoplasmic and nuclear protein determination following permeabilization of the cells
- Detection of rare cell populations such as stem cells, progenitors and immunospecific reactive cells
Theory and practice
Excellent multimedia presentations on the theory and practice of flow cytometry:
A comprehensive text on the chemistry and use of fluorescent probes:
Resources
Spectral Viewers
Equipment and facilities
Flow Cytometers
The Flow Cytometry Core Facility has two multiuser benchtop flow cytometers located within the School Of Medicine, Ninewells Hospital. They are situated within The Immunoassay Biomarker Core Laboratory, L7/181.
The Becton Dickinson LSRFortessa is a benchtop analyser equipped with three lasers, a 488nm, a 633nm and a 405nm, which allows up to thirteen parameter analysis (two scatter detectors and eleven colour specific detectors).
The Attune Acoustic Focusing flow cytometer is equipped with 405nm and 488nm lasers and capable of 8 parameter analysis (two scatter detectors and six colour specific detectors). This system also contains an Autosampler allowing uptake of samples from multiwell plates.
In general, the analysers are capable of detecting any fluorochrome that is excitable by the laser(s) contained in that analyser.
In addition to the software specific to these machines, registered users have access to the analysis package Flowjo on a standalone analysis platform.
Time on a benchtop analyser may be booked by registered staff and students using our online booking system.