Submitted by PRACHEE RAJPUT

Student of M.Sc sem –II

DEPT OF ZOOLOGY AND APPLIED

AQUACULTURE B.U BHOPAL

SYNOPSIS

• INTRODUCTION• DEFINITION• APPLICTION• ACTION & WORKING• CONLUSION• REFERENCES

What cells are in there?

How can we use Flow Cytometry to quantify cells?

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

• Clinical

•Research

Dr. Allan MCB Harvard

NeutrophilsLymphocytesMonocytesEosinophilsBasophils

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

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?

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/cbcwbc.html

Why do we need technology?

How it works1. 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

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.

NeutrophilsLymphocytesMonocytesEosinophilsBasophils

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 CellOrange Runt NeutrophilGreen Runt MonocyteYellow Runt BasophilPink Runt EosinophilWhite Nerds Lymphocyte

What can this differential tell you?

http://www.bioteach.ubc.ca/MolecularBiology/FlowCytometry/

See real Data

Let’s take a closer look.

How can we differentiate T cells?

http://www.flow-cytometry.de/

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 cellsPink Nerds += CD4+ Helper T cells

Categorize and count your T cell population

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

REFERENCES

• INSTRUMENTAL BIOLOGY• INTERNET SOURCES