Simultaneous Measurement of Necrotic,

1
Simultaneous Measurement of Necrotic,
Apoptotic Dead and Live Cells
Necrotic/Apoptotic Dead
Treat Cells with DNAse to nick cells
Incorporate FITC-UTP with TdT
Apoptotic Live
Fix cells and permeabilize
Incorporate B-UTP with TdT then
PE-Avidin
2
SIMULTANEOUS EVALUATION OF DEAD, NECROTIC AND
APOPTOTIC CELLS
0
1
2
3
4
10
10
10
10
10
0
256
512
768
1024
FITC UTP -gt
FSC -gt
TDT48015
TDT48015
3
LIGHT SCATTER ANALYSIS OF DEAD CELLS AND VIABLE
APOPTOTIC CELLS
R4
R3
gated R1
A
B
PE-dUTP
SSC
R1
R2
Fl-12-dUTP
FSC
gated R2
gated R3
C
D
SSC
SSC
FSC
FSC
4
FOUR COLOR DETECTION OF SURFACE ANTIGENS,
APOPTOSIS AND DEAD CELLS
DEAD CELLS
PE-CD19
DEAD CELLS
APOPTOSIS
TC-CD33
APOPTOSIS