Biology Reference
In-Depth Information
4. A stable baseline is obtained quickly only if the dye released from cells by
digitonin permeabilization is swept rapidly away from the region directly above the
microscope objective. Otherwise, fluorescence from the released dye still will be
captured by the objective and, thus, contribute to the measured background. Once
swept away and diluted into the bulk medium, the released dye contributes
negligibly to the background.
A
45 6
123
F
340
F
f,380
F
380
BG
340
BG
380
F
b,380
0
20
40
60
Time (min)
80
100
120
B
20
5
R
max
0.8
13
2
0.7
10
6
0.6
0.5
20
40
60
80
100
R
min
0
0
20
40
60
Time (min)
80
100
120
Fig.11
(Continued)
