Isolation of Lymphocytes

Figure A-23

Isolation of peripheral blood lymphocytes by FicollHypaque gradient

11

Stimulaton of Lymphocytes

Figure A-35

22

Flow cytometry
Definition:
a

technique wich cells/ events passed

individually through a beam of laser light,
amplified and converted to digital signal and
can be plotted to form a scattergram.
Cells or other particles can be analyzed using
fluorescence technique resulted after
previously fluorochrome labeled

33

flow cytometry technology

Characterize cells:
Individually
Fast
Multiparametric
Quantitatively

Specific
sub population

diagnostic &
prognostic
information

Clinical
application

44

Cells analysis using FACS

subset
receptore
n

activatie

cel cyclus

adhesie

apoptose

chemokin
e
cytokine
55

Distribution of lymphocyte subpopulation in human

peripheral blood

Figure A-24

66

Subset limfosit
B

CD19+
T-helper

CD3+ CD4+
T-cyt.

CD3+ CD8+
77

1
88

99

Flow cytometry and FACS

analysis

Figure A-25 part 1 of 2

1010

Forward Scatter (FSC) detection

Laser

FSC
detector

1111

Side Scatter (SSC) detection

Laser

FSC detector
90o

SSC detector
1212

Fluorescence detection

FSC detector

Laser
90o

Fluorescence detector
1313

Multiparameter Assessment

1414

1515