Flow Cytometry Applications

Flow cytometric applications available at the Malaghan Institute:

1.     Cell surface expression (immunophenotyping)
  • For example flow cytometry is currently the most sensitive technique for being able to characterise effector and memory T cell responses to vaccines.  The LSRII now enables our researchers to investigate and define the full magnitude and quality of the T cell response by monitoring CD4+, CD8+ and T regulatory cells simultaneously, and looking at their ability to proliferate or induce proliferation (growth factor secretion), terminate, activate, or organise the immune response (cytokine/chemokine production).  The LSRII can also be used to determine whether vaccines are inducing a memory or effector T cell population.         

2.     Absolute cell counts

3.     Cell type enumeration

4.     Intracellular antigen expression (cytokine/chemokine production)

5.     DNA analysis (cell cycle analysis)

  • Using standard DNA binding dyes (for example, Hoechst, DAPI, PI, 7-AAD)

6.     Protein expression and localisation

7.     Mitochondrial membrane potential (redox state of cells)

8.     Calcium flux measurement 

  • Using Indo-1

9.     Morphological complexity (gross cellular changes)

10.   Cellular kinetics (proliferation)

11.   Viability

12.   Apoptosis

13.   Cell sorting 

  • We have the capability to do: sterile cell sorting, providing chilling for the sample and collected cells; sorting of stem cell side populations using Hoechst 33342; sorting of up to four populations of cells simultaneously into Eppendorf or 5 mL tubes; sorting (up to 7 colours) and analysis (up to 18 colours) of multicolour immunophenotyped samples; sorting and analysis of CFP, GFP, YFP, mRFP, mTomato and mCherry gene expression proteins

14.   Consulting on experimental design and effective flow cytometry components for grants

 

These resources are made equally available to Malaghan Institute researchers, as well as to researchers within other academic and commercial entities.

For more information, please contact:

Kylie Price
Flow Cytometry Suite Manager
Tel: +64 4 499 6914
Email: kprice@malaghan.org.nz