Flow cytometry is a technique that analyzes physical and chemical characteristics of cells as they flow in a fluid stream through a beam of light. It is used for immunophenotyping, DNA and blood analysis, and diagnosis of health disorders. The main components are fluidics to transport cells, optics for excitation and detection of light, and electronics to convert light signals into data. Gating is used to select subsets of cells for further analysis. Flow cytometry has applications in clinical settings and research, such as monitoring HIV, identifying lymphocyte subsets, and diagnosing hematological cancers.

1. Andrea Ajero
Solomon Ihejinka
Mayur Patel
Diana Asumang
Nebiha Ibrahim
Renee Backen

2. DEFINITION OF FLOW CYTOMETRY?
 Flow - motion Cyto - cell Metry - measure
FACS (Flourescence Activated Cell Sorting)
 PURPOSE S OF FLOW CYTOMETRY
 Immunophenotyping
 DNA and Blood Analysis
 Data Analysis
 Diagnosis (Health Disorder)
THE PRINCIPLES OF FLOW CYTOMETRY
THE APPLICATIONS OF THE METHODOLOGY IN THE CLINICAL
LABORATORY
 Clinical
 Research

3. PRINCIPLE
 -Light source
 -Lasers
 Light scattering/Reflection
 -FSC (Forward scatter)
 -SSC (Side scatter)
 -Light excitation
 -Wavelength
 -Fluorophores emission
 -Recognition of cell
 -Histogram
 -One parameter
 -Two parameter

4. PRINCIPLE
• The three main subsystems-:
- Fluidics – It transport particles to the laser beam for interrogation.
- Optics – Excitation and Collection Components.
- Electronics – It convert detect lights into signals and send information to
computer. The data is stored and graphs are plotted to further analysis it.

5. The process of gating is very important
in the analysis of flow cytometry data
Gating refers to the process of selecting
a subset of the collected events for
further analysis
Subsets can continue to be gated to
generate further subsets
The process of gating simply creates a
new population
Naming subpopulations in gating is important
 Siblings with the same names are not allowed,
otherwise there would be confusion
It is recommended that using duplicate names of
subpopulations within any given sample be avoided
Gating

6. WHERE CAN FLOW CYTOMETRY APPLICATION BE
FOUND?
Flow Cytometry Facilities
 Mayo Clinic
 Stanford School of Medicine
 Unc. Sick Kids Research Clinic
in Canada
 Veterinary Services
 George Washington University
Medical Center
 The University of Chicago
 John Hopkins Medical
Research
Mayo Clinic Locations:
 Minnesota , Arizona,
Wisconsin,
 Georgia, Arizona, Florida,
Massachusetts
 Amyloidosis
Flow cytometric immunophenotyping showed
an abnormal B-cell population expressing
CD19, CD20, dim monotypic lambda light
chains, CD23, CD5, consistent with CLL.

7. Flow Cytometer’s
Application in Clinical
And Research
Monitoring HIV—Identification and
Enumeration
of Lymphocyte Subsets
The BD Multitest™ 6-color TBNK kit is the first
and only 6-color In Vitro Diagnostic application
to provide a complete immune panel in a single
tube, saving valuable time and resources for
sample processing

8. References
BD Bioscience (2011) BD FACSCantoTM Brochure. Retrieved from
http://www.bdbiosciences.com/documents/BD_FACSCanto_II_brochure.pdf
Brown, M., & Wittwer, C. (2000, August). Flow Cytometry:
Principles and Clinical Applications in Hematology.
Retrieved September 15, 2013 from
http://www.clinchem.org/content/46/8/1221.full
Clinical Flow Wiki. (n.d.) Introduction to Flow Cytometric Analysis Retrieved from
http://www.clinicalflow.com/Cases/Introduction_to_Flow_Cytometric_Analy
sis
Evans, F. (2012). Fiber delivery systems enhance flow cytometry designs. Laser
Focus World, 48(11), 53. Retrieved from
http://web.ebscohost.com/ehost/pdfviewer/pdfviewer?sid=378fd7c5-f697-
49d2-88cf-e51f160b7b9b%40sessionmgr4&vid=4&hid=9

9. References
FlowJo - v7.6.5 Manual - Gating."FlowJo Home”. N.p. (n.d.) Retrieved from
http://www.flowjo.com/v765/en/gating.htm
Fromm, J., Thomas, B., Wood, B. (2009) Flow Cytometry Can Diagnose Classical
Hodgkin Lymphoma in Lymph Nodes with High Sensitivity and Specificity
American Journal of Clinical Pathology. Retrieved from
http://www.medscape.com/viewarticle/453271
Introduction to Flow Cytometry: A Learning Guide, (n.d.) “Stem Cell Institute Medical
School. Version 11-11032-01. University of Minnesota. Retrieved from
http://www.stemcell.umn.edu/prod/groups/med/@pub/@med/documents/as
set/med_80691.pdf
Kincaid, K. Semiconductor lasers shed new light on flow cytometry (2004). Laser
Focus World. Retrieved from
http://web.ebscohost.com/ehost/pdfviewer/pdfviewer?sid=378fd7c5-f697-
49d2-88cf-e51f160b7b9b%40sessionmgr4&vid=4&hid=9

10. References
Lookfordiagnosis (n.d.) Flow Cytometry. Retrieved from
http://www.lookfordiagnosis.com/mesh_info.php?term=Flow+Cytometry
&lang=1
Mayo Foundation for Medical Education and Research. (1995-2013) Flow
Cytometry Immunonphenotyping. Retrieved from
http://www.mayomedicallaboratories.com/articles/hottopics/transcripts/
2009/2009-12a-amyloidosis/12a-17.html
Overview-BD (n.d.) Medical Supplies, Devices and Technology. Retrieved from
http://www.bd.com/videos/bdb/training/itf/overview/player.html
Science Art. (2008). Retrieved September 15, 2013, from
http://vesuvias.wordpress.com/page/2/
Shen, H., Tang, Y. (2012) Rapid Detection of Neoplastic Cells in Serous Cavity
Effusions in Children with Flow Cytometry Immunophenotyping. Retrieved
from http://www.ncbi.nlm.nih.gov/pubmed/22288605

11. References
Science Art. (2008). Retrieved September 15, 2013, from
http://vesuvias.wordpress.com/page/2/
Shen, H., Tang, Y. (2012) Rapid Detection of Neoplastic Cells in Serous
Cavity Effusions in Children with Flow Cytometry Immunophenotyping.
Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/22288605