1. Introduction To Flow Cytometry:
Niga Nawroly

2. What is flow cytometry?
Flow cytometry is a method of measuring multiple physical
and chemical characteristics of particles by optical means.
Peripheral blood, Bone marrow cells
Bacteria
Yeast

3. Applications of Flow Cytometry.
Cell size.
Cytoplasmic granularity.
Cell surface antigens (phenotyping).
Apoptosis.
Intracellular cytokine production.
Intracellular signalling.
Gene reporter (GFP).
Cell cycle, DNA content, composition, synthesis.
Bound and free calcium.
Cell proliferation (BRDU and CFSE)
Cell sorting, single cell cloning (clonecyt)

4. + Phenotyping, Size and granularity detection: Sample requirements:
A suspension of single cells or other particles
in a suitable buffer, usually PBS.
Typical density : cells / ml +
Incubate
Acquire

5. A cytometer consists of:
Electronics
Fluidics
Light detectors
FL3 PerCP, Cy5
Computer
FL2 PE
FL1 FITC
SSC
Laser
Light
488nm
FSC

6. Size and Granularity Granularity Size All Data shown is from:
BAL from Balb/c primed with TT
Data produced by Dr John Tregoning

7. Presenting and Interpreting Data
Single parameters can be displayed as a histogram.
Dual parameter data can be displayed in two dimensions using dot, density or contour plots.

8. Gating:

9. Back-gating:

10. Available analysis software
Cellquest
WinList
Flowjo
WinMDI
Free cytometry software:

11. Flow Cytometry : Reference Material
Books
Practical flow Cytometry Howard M Shapiro.
Flow Cytometry: A Practical Approach MG Ormerod.
Introduction to Flow Cytometry, JV Watson.
Flow Cytometry: First Principals, Alice Givan.
Cytometric Analysis of Cell Phenotype and Function, McCarthy & Macey.
Journal
Cytometry : Journal of the International Society for Analytical Cytology.

12. Flow Cytometry on the web
ISAC (International Society For Analytical Cytology)
Salk Institute
Purdue University
Scripps Research Institute
Cancer Research UK

13. Our website New Website address: http://www.wfi.med.ic.ac.uk/Flow.htm