Transcript FACS flourescens activeted cell sortering

FACS
Flourescens Activeted Cell sortering
användningsområden
FACS diagram
FACS modell
Hydrodynamisk fokusering
laminärntflöde
Tubulent flöde
Laminärtflöde parametrar
Skum
Foam
The relatively stable foam of beer, for
example, results from an interaction
between a protein fraction rich in
basic and aromatic amino acids and
the hop bitter compounds. Developing
and improving knowledge of the
relationship between peptide
concentration and structure and foam
performance can lead to improved
understanding, analysis and control of
this complicated property of food
products.
Dr. Karl J. Siebert - (Tel: 315-787-2299) Email:
[email protected]
Ljusspridning - partikelstorlek
Scattering
Forward scatter
Side scatter
http://www.epa.gov/owow/volunteer/proceedings/sixth/session1.html
Side scatter vs Forward scatter
University of Utah
scatterplot
fluorimeteroptik
Vaccine Research Center / 40 Convent Drive / Bethesda, Maryland 20892
Fluorimeter
Flourescens färger
Fluorescein
Fluorescens plott
Lin och log
Sample : peripheral blood after red cell lysis.
CD14 PE (log)
Side Scatter (linear)
Data collected on 10,000 cells.
CD45 FITC (log)
Forward Scatter (linear)
R1=monocytes (CD14+ve)
R2=lymphocytes (CD45>monocytes)
R3=granulocytes (CD45<monocytes)
[email protected]
Isometric Displays
Sample : peripheral blood after red cell lysis.
Data collected on 10,000
Events
High
Events
CD14 PE
Low
CD 45 FITC
Scatter
Fluorescence
[email protected]
Apoptotiska celler
As cells die or become apoptotic the refractive index of the internal cytoplasm
becomes more similar to that of the extracellular medium - this manifests
itself as a reduction in forward scatter signal. At the same time, intracellular
changes and invagination of the cytoplasmic membrane lead to an increase
in side (or orthogonal or 90°) scatter. If we add a dead cell discriminatory dye
we can identify cells that have become permeable. In this way we can get
low level resolution of dead and apoptotic cells.
science.cancerresearchuk.org/.../images/51998
endotelceller
R2= total cells
R1= living cells
Isolation of lymph endothelial cells (LEC) from dermal skin capillary ECs
a case study with cells from one donor pre-selected by CD31 magneto bead
sorting
www.reliatech.de