Flow cytometry is a process that can quantify and categorize cells by measuring their physical and chemical characteristics. Cells pass through a flow cytometer one by one where they are hit by a laser beam, scattering light. Detectors then measure the light and determine characteristics like size, shape, and protein expression. This allows cells to be separated into subpopulations and quantified. Flow cytometry is useful for clinical and research applications like performing complete blood counts and identifying T cell subtypes.

1. Submitted by
PRACHEE RAJPUT
Student of M.Sc sem –II
DEPT OF ZOOLOGY AND
APPLIED
AQUACULTURE
B.U BHOPAL

2. SYNOPSIS
• INTRODUCTION
• DEFINITION
• APPLICTION
• ACTION & WORKING
• CONLUSION
• REFERENCES

3. What cells are in there?
How can we use Flow Cytometry to
quantify cells?

4. What is flow Cytometry?
Process for quantifying cells
•Measures different property of cells
•Able to categorize and quantify
•Even able to separate out
subpopulations of cells

5. • Clinical
•Research
Dr. Allan MCB Harvard

6. Neutrophils
Lymphocytes
Monocytes
Eosinophils
Basophils
White Blood Cells,
Platelets (stained
purple), a T-
Lymphocyte white
cell (stained
green), and a
Monocyte white
cell (stained gold)
as seen through a
scanning electron
microscope.
©2000 Dennis
Kunkel, Ph.D.
Image Source: http://www.mybloodyourblood.org/biology_white.htm

7. How could you separate your sample
of white blood cells into
subpopulations?
What characteristics of the cell
could you use?
How about if they had no color?

8. Standard Differentials
1. Draw sample from patient
2. Smear blood on slide
3. Stain with differential WRIGHT
stain
4. Count (at least 100) http://www.rnceus.com/cbc/cb

9. Why do we need technology?

10. How it works
1. Draw cells, with excess fluid, from test tube into machine.
2. Cells pass in single file past laser.
3. Laser hits cell and light is scattered.
4. Photomultiplier multiplies light intensity and a light sensor measures the
amount of light and scatter pattern.
5. Based on cell characteristics (size and shape), the computer categorizes and
quantifies the cells.
Click to learn more

11. Create a table to record data for your white
blood cell differential.
** Remember you will need to record the
number of each cell in your sample.
Neutrophils
Lymphocytes
Monocytes
Eosinophils
Basophils

12. Use the following key to categorize
your sample of White Blood Cells.
What is the percent break up of
your white blood cell differential?
Candy Cell
Orange Runt Neutrophil
Green Runt Monocyte
Yellow Runt Basophil
Pink Runt Eosinophil
White Nerds Lymphocyte

13. What can this differential tell you?
http://www.bioteach.ubc.ca/MolecularBiology/FlowCytometry/
See real Data

14. Let’s take a closer look.
How can we differentiate T cells?
http://www.flow-cytometry.de/

15. Create a table to categorize and count your
T-cell subpopulation into CD4+ Helper T cells
and CD8+ Cytotoxic T cells.
Purple Nerds= CD8+ Cytotoxic T cells
Pink Nerds += CD4+ Helper T cells
Categorize and count your T cell population

16. CONCLUSION
• From the above discussion we have
concluded that flow cytometry is helpful to
stud the property of cell
• It aslo helpful in clinical and research

17. REFERENCES
• INSTRUMENTAL BIOLOGY
• INTERNET SOURCES