Flow cytometry
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Flow cytometry. Powerful technique for analysis of multiple parameters of individual cells within heterogeneous populations multiple physical characteristics of single particles or cells. What is flow cytometry?. scheme of flow cytometry. The fluidic system & optics system.

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PowerPoint Slideshow about 'Flow cytometry' - Renfred


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What is flow cytometry l.jpg

Powerful technique for

analysis of multiple parameters of individual cells within heterogeneous populations

multiple physical characteristics of single particles or cells

What is flow cytometry?



The fluidic system optics system l.jpg
The fluidic system & optics system

  • heart of the system

  • where the laser and the sample intersect and the optics collect the resulting scatter and fluorescence


The fluidic system l.jpg
The fluidic system

  • Particles or cells are passed through the laser beam one at a time

Laser

Hydrodynamic focusing


Size comparison by flow cytometry l.jpg
Size comparison by flow cytometry

  • ~ 1 ~ 15 uM size detection


Forward scatter l.jpg
Forward scatter

  • Light is quantified by a detector that converts intensity into voltage

  • Cell size


Side scatter l.jpg
Side scatter

  • Granularity and structural complexity inside the cell


Scatter plot l.jpg
Scatter plot

SSC

FSC






Two color experiment14 l.jpg
Two color experiment

-/+

+/+

-/-

+/-



Compensation16 l.jpg
Compensation

  • No-stained sample

  • Only FITC stained sample

  • Only PE stained sample

uncompensated

compensated

PE

FITC


Application apoptosis l.jpg
Application (apoptosis)

  • Apoptosis (FSC/SSC)

Apoptosis

SSC

Necrosis

FSC


Application apoptosis18 l.jpg
Application (apoptosis)

  • Apoptosis (Annexin V/PI assay)

Normal Cell

Apoptotic Cell

: Phosphatidyl Serine

: Annexin V-FITC


Application apoptosis19 l.jpg
Application (apoptosis)

  • Apoptosis (Annexin V/PI assay)

Control

Anti-Fas Ab

Propidium Iodide (PI)

Annexin V-FITC


Application cell cycle l.jpg
Application (cell cycle)

Apoptotic DNA Histogram

A typical DNA Histogram

G0-G1

G2-M

S

Apoptotic cells

Normal

G0/G1 cells

# of Events

Fluorescence Intensity


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